FFPE RNA/DNA Purification Kit

FFPE RNA/DNA Purification Kit

Catalog# : NB-25000
Price/Unit : $295.00
Unit : 50 preps

For the rapid isolation and purification of total RNA (including microRNA) and genomic DNA from FFPE tissue samples.

* Purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA)
* Preferential purification of RNA from other cellular components without the use of phenol or chloroform
* Purified RNA and genomic DNA are of the highest quality for use in numerous downstream applications
* Available in 96 well format

Bio-Synthesis's FFPE RNA/DNA Purification Kit provides a rapid method for the isolation and purification of total RNA (including microRNA) and genomic DNA from formalin-fixed paraffin-embedded (FFPE) tissue samples. Alternatively, the kit can be used to isolate total RNA alone or genomic DNA alone from FFPE tissue samples through varying the protease digestion time and performing optional RNase or DNAse digestions.

Using formalin to fix tissues leads to crosslinking of the nucleic acids and proteins, and the process of embedding the tissue samples can also lead to fragmentation of the nucleic acids over time. Bio-Synthesis's FFPE RNA/DNA Purification Kit provides conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of nucleic acids.

Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. The process first involves deparaffinization of the FFPE samples through a series of xylene and ethanol washes. Next, the FFPE samples are digested with the provided Proteinase K and Digestion Buffer. Binding Solution and ethanol are then added to the lysate, and the solution is loaded onto a spin-column (BIND). Under these conditions only the RNA and/or DNA will bind while most of the contaminants will be removed in the flowthrough. At this point, any traces of genomic DNA can be digested allowing for pure RNA samples to be isolated. Alternatively, traces of the RNA may be digested at this point if a pure sample of genomic DNA is required instead. The bound nucleic acid is then washed (WASH) in order to remove any impurities. Lastly, the purified total nucleic acd is eluted in 50 µL of the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart to the right.

This kit is also available in a 96-well format for high-throughput FFPE RNA/DNA purification. Purifictaion with the 96-well plate can be performed using either a vacuum manifold or centrifugation.
FEATURES AND BENEFITS

* High quality and integrity of the isolated nucleic acid - Both the purified total RNA and/or genomic DNA are of the highest quality and integrity, and can be used in any sensitive downstream application.
* Isolate a diversity of RNA species - All RNA species can be isolated, from large mRNA and ribosomal RNA down to microRNA and siRNA.
* High yields - Bio-Synthesis's FFPE RNA/DNA Purification Kit allows for the purification of high yields of total RNA or DNA.
* Rapid procedure - Isolate total RNA and/or genomic DNA from FFPE tissue sections using a rapid spin column format or a high-throughput 96-well plate format.
* Versatile procedure - The kit allows for the purification of either pure total RNA, pure genomic DNA or both RNA and genomic DNA through the use of tailored protease digestion times.

APPLICATIONS

The purified RNA and genomic DNA are of the highest available quality from FFPE samples, and can be used in a number of downstream applications including:

* qPCR
* qRT-PCR
* Reverse transcription PCR
* Primer extension
* Mutation screening
* Expression array assays
* Microarray analyses
* Sequencing
* Southern blotting
* SNP analysis
* Next Generation Sequencing
Figure 1. High Quality and Yield of Total RNA. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Bio-Synthesis's FFPE RNA/DNA Purification Kit and a leading competitor’s kit. One microliter of the 50 µL purified RNA was then resolved on an Agilent 2100 BioAnalzyer using an RNA Nano 6000 chip. As it can be seen, Bio-Synthesis's not only isolated higher yields of total RNA, but the RNA was also of a higher quality as evidenced by the higher RIN values obtained with Bio-Synthesis's RNA.

FFPE RNA/DNA Purification Kit Contents
1. Digestion Buffer
2. Binding Solution
3. Enzyme Incubation Buffer
4. Wash Solution
5. Elution Buffer
6. Proteinase K
7. Spin Columns
8. Collection Tubes
9. Elution Tubes
10. Product Insert
Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. The Proteinase K should be stored in aliquots at -20°C upon reconstitution. These reagents should remain stable for at least 1 year in their unopened containers.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.

FFPE RNA Purification Kit

FFPE RNA Purification Kit

Catalog# : NB-25300
Price/Unit : $320.00
Unit : 50 preps

For the rapid and efficient extraction and purification of RNA (including microRNA) from FFPE samples

* Extract total RNA (including microRNA) from FFPE samples
* No phenol extraction step
* Includes DNAse for optional on-column DNA removal
* Isolated RNA is of the highest quality and integrity

Bio-Synthesis's FFPE RNA Purification Kit provides a rapid method for the isolation and purification of total RNA (including microRNA) from formalin-fixed paraffin-embedded (FFPE) tissue samples in as little as 1 hour. Using formalin to fix tissues leads to crosslinking of the RNA and proteins, and the process of embedding the tissue samples can also lead to fragmentation of the RNA over time. Bio-Synthesis's FFPE RNA Purification Kit provides conditions that allow for the partial reversing of the formalin modifications, resulting in a high quality and yield of RNA. The kit is able to purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA), depending on the age of the FFPE tissue as fragmentation of the RNA is known to occur over time.

Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. The process first involves deparaffinization of the FFPE samples through a series of xylene and ethanol washes. Next, the FFPE samples are digested with the provided Proteinase K and Digestion Buffer. Binding Solution and ethanol are then added to the lysate, and the solution is loaded onto a spin-column (BIND). Under these conditions only the RNA will bind while most of the contaminants will be removed in the flowthrough. At this point, any remaining traces of genomic DNA can be digested using an optional protocol, allowing for pure RNA samples to be isolated. The bound RNA is then washed (WASH) in order to remove any impurities. Lastly, the purified total RNA is eluted in 50 µL of the provided Elution Buffer or water (ELUTE). Please see the procedure flowchart to the right.

FEATURES AND BENEFITS

* High quality and integrity of the isolated RNA - The purified total RNA is of the highest quality and integrity, and can be used in any sensitive downstream applications.
* Isolate a diversity of RNA species - All RNA species can be isolated, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA).
* High yields - Bio-Synthesis's FFPE RNA Purification Kit allows for the purification of high yields of total RNA.
* No phenol:chloroform extractions - Total RNA is isolated from FFPE tissue samples without the use of harmful chemicals such as phenol or chloroform.
* Rapid procedure - Isolate total RNA from FFPE tissue sections using a rapid spin column format in as little as 1 hour

APPLICATIONS

Purified RNA is of the highest quality, and can be used in a number of downstream applications including:

* Bioanalyzer
* Quantitative, real-time RT-PCR for both large mRNA and small RNA including miRNA
* RT-PCR for both large mRNA and small RNA including miRNA
* Northern blotting
* RNase protection
* Primer extension
* Expression array assays
* Next Generation Sequencing
* microRNA Cloning

Figure 1. High Quality and Yield of Total RNA. Bio-Synthesis's FFPE RNA Purification Kit isolates FFPE RNA that exceeds both yield and quality of competitors. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Bio-Synthesis's FFPE RNA Purification Kit and a leading competitor’s kit. One microliter of the 50 µL purified RNA was then resolved on an Agilent 2100 BioAnalzyer using an RNA Nano 6000 chip. As it can be seen, Bio-Synthesis's not only isolated higher yields of total RNA, but the RNA was also of a higher quality as evidenced by the higher RIN values obtained with Bio-Synthesis's RNA.

Figure 2. Higher Yield of FFPE RNA Isolated by Bio-Synthesis's FFPE RNA Purification Kit. Bio-Synthesis's FFPE RNA Purification Kit isolates FFPE RNA that exceeds the yield of competitors. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Bio-Synthesis's FFPE RNA Purification Kit and two leading competitor’s kits. Eighteen isolations were performed for each product. The graph demonstrates the mean yield of RNA according to spectrophotometry for 18 sample replicates. Vertical bars represent the standard deviation. Bio-Synthesis's kit consistently purified total RNA with a higher yield than for those obtained using the market competitor's kits.

Figure 3. High Yield of a Diversity of RNA Species. Bio-Synthesis's FFPE RNA Purification Kit effectively recovers all sizes of RNA from large mRNA to small RNA including microRNAs. Total RNA was isolated from one slice of hamster FFPE kidney section (20 micron thickness) using Bio-Synthesis's FFPE RNA Purification Kit and a leading competitor’s kit. Five microliters of the 50 µL purified RNA was then used as the template in a 20 µL reverse transcription reaction with oligo dT primer or miR-21 stem-loop reverse primer. Two microliters of the RT reaction was then used in a 15 µL qPCR reaction for detecting the beta-actin gene (Panel A) and for detecting miR-21(Panel B), respectively. In both graphs the blue lines correspond to Bio-Synthesis's isolated-RNA and the green lines correspond to competitor-isolated RNA. As it can be seen, Bio-Synthesis's kit isolated higher yields of RNA in both cases, as indicated by the lower Ct values of the blue lines. Also, Bio-Synthesis's kit successfully isolated not only large RNA (Panel A) but also microRNA (Panel B), indicating the diversity of RNA species isolated.

FFPE RNA Purification Kit Contents - Spin Columns
1. Digestion Buffer
2. Binding Solution
3. Enzyme Incubation Buffer
4. Wash Solution
5. RNA Elution Buffer
6. Proteinase K
7. DNase I
8. Spin Columns
9. Collection Tubes
10. Elution Tubes
11. Product Insert
Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. The DNAse I should be stored at -20°C upon arrival. The Proteinase K should be stored in aliquots at -20oC upon reconstitution. These reagents should remain stable for at least 1 year in their unopened containers.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.

RNA Clean-up and Concentration Kit

RNA Clean-up and Concentration Kit

Catalog# : NB-23600
Price/Unit : $240.00
Unit : 50 preps
For rapid and efficient clean-up and concentration of total RNA, including microRNA, without phenol

* Clean-up and concentrate total RNA in minutes
* No phenol or chloroform extractions
* Clean-up RNA from contaminants including enzymes, RNases and nucleotides
* Rapid spin-column protocol
* Purified RNA is fully compatible with all downstream applications

Bio-Synthesis's RNA Clean-Up and Concentration Kit provides a rapid method for the purification, cleanup and concentration of up to 35 μg of RNA isolated using different methods including phenol/guanidine-based protocols and from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. The kit can concentrate as much as 35 µg of RNA to as little as picograms of RNA. The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA). The RNA is preferentially purified from other reaction components such as proteins, RNases and nucleotides without the use of phenol or chloroform.

Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. Briefly, the RNA samples or reactions containing RNA are mixed with Binding Solution, ethanol is added and the RNA is bound to Bio-Synthesis's column (BIND). Under these conditions only the RNA will bind to Bio-Synthesis's resin while most of the contaminating proteins and nucleotides are removed in the flowthrough. The bound RNA is then washed to remove any remaining impurities (WASH). Lastly, the purified total RNA is eluted into 20 - 50 µL of the provided Elution Buffer or water (ELUTE). Please see procedure flowchart to the right.

The purified RNA is of the highest integrity, and can be used in a number of downstream applications. Bio-Synthesis's RNA Clean-Up and Concentration Kit purifies RNA with minimal amounts of DNA contamination. An optional protocol is provided for maximum removal of residual DNA that may affect sensitive downstream applications such quantitative PCR.


FEATURES AND BENEFITS

* Efficient clean-up from various enzymatic reactions - RNA clean-up from upstream enzymatic applications such as labeling, DNase treatment and in vitro transcription. Remove proteins, RNases, DNases, nucleotides, etc.
* Clean-up and concentration of RNA isolated by different methods - RNA that has been isolated using various methods, including phenol-chloroform extractions or alcohol precipitations, can be processed with this kit
* Fast and easy processing - Rapid spin-column format allows for the processing of 10 samples in 20 minutes
* No phenol:chloroform extractions - Bio-Synthesis's RNA Clean-Up and Concentration Kit purifies RNA without the use of harmful chemicals such as phenol or chloroform.
* Isolate all small RNA molecules - All small RNA species can be isolated including microRNA, siRNA, tRNA, and 5S rRNA
* Recovered RNA is suitable for downstream applications - Purified RNA can be used in a number of downstream applications including end-point or quantitative reverse transcription PCR, Northern blotting, and expression array analysis


APPLICATIONS
Purified RNA is of the highest quality and can be used in a number of downstream applciations including:

* Bioanalyzer
* Quantitative, real-time RT-PCR for both large mRNA and small RNA including miRNA
* RT-PCR for both large mRNA and small RNA including miRNA
* Northern blotting
* RNase protection
* Primer extension
* Expression array assays
* Next Generation Sequencing for RNA and miRNA
* microRNA Cloning

Figure 1. Effective Clean-Up to Produce High Quality Total RNA with Complete Size Diversity. Bio-Synthesis's RNA Clean-Up and Concentration Kit effectively cleans up RNA isolated from phenol-based extractions without the loss of RNA diversity by retaining all RNA species including small RNAs. Total RNA was isolated from 5 x 108 E. coli using a competitor’s phenol-based RNA extraction reagent. The resulting RNA was then purified using Bio-Synthesis's RNA Clean-Up and Concentration Kit. As controls, total RNA was extracted using both Bio-Synthesis's Total RNA Purfication Kit (#17200, no phenol required) and the phenol-based RNA extraction reagent only without any clean-up. Resolution of 7 µL of the 50 µL purified RNA on a 1X MOPS, 1.5% formaldehyde-agarose gel showed the RNA extracted with phenol was successfully cleaned-up by Bio-Synthesis's RNA Clean-Up and Concentration Kit without the loss of small RNA species.
Figure 2. Effective Clean-Up to Produce High Quality Total RNA Compatible to Bioanalyzer Analysis. Total RNA was isolated from 0.75 million HeLa cells using a competitor’s phenol-based RNA extraction reagent. The resulting RNA was then purified using Bio-Synthesis's RNA Clean-Up and Concentration Kit. As controls, total RNA was extracted using both Bio-Synthesis's Total RNA Purfication Kit (#17200, no phenol required) or the phenol-based RNA extraction reagent only without clean-up. Resolution of 1 µL of the 50 µL purified RNA on an Agilent RNA Nano 6000 Chip showed that without clean-up, the RNA sample isolated using the phenol-based RNA extraction reagent only resolved poorly on a bioanalyzer. In contrast, RNA extracted with the phenol-based RNA extraction reagent followed by clean-up by Bio-Synthesis's RNA Clean-Up and Concentration Kit resolved properly on the bioanalyzer showing excellent quality.

RNA Clean-Up and Concentration Kit Contents
1. Spin Column
2. Binding Solution
3. Wash Solution
4. Elution Buffer
5. Micro Spin Columns
6. Collection Tubes
7. Elution Tubes
8. Product Insert
Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.

CleanAll RNA/DNA Clean-Up and Concentration Kit

CleanAll RNA/DNA Clean-Up and Concentration Kit

Catalog# : NB-23800
Price/Unit : $240.00
Unit : 20 preps
For the rapid and efficient purification, cleanup and concentration of RNA or DNA

* Purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA)
* RNA is preferentially purified from other reaction components such as proteins and nucleotides, without the use of phenol or chloroform
* Purifies all sizes of DNA, from small PCR products to plasmids to genomic DNA.
* Rapid and efficient spin-column format

Features and Benefits | Specifications

BSI's CleanAll RNA/DNA Clean-Up and Concentration Micro Kit provides a rapid method for the purification, cleanup and concentration of RNA or DNA from different isolation methods or upstream applications. The kit can be used as an alternative to organic extraction and ethanol precipitation to clean up various enzymatic reactions. The CleanAll Kit purifies RNA from phenol/guanidine-based protocols or from various upstream enzymatic reactions such as DNase treatment, labeling and in vitro transcription. Furthermore, endotoxins can be removed from previously purified RNA solutions. The kit can be used to clean up DNA from digestions, ligations, PCR reactions, labeling reactions, DNA modification reactions and staining. The kit also provides a protocol for the rapid removal of endotoxins from previously purified DNA down to 0.1 EU/µg DNA or less.
Purification is based on spin column chromatography using BSI’s proprietary resin as the separation matrix. Briefly, the samples or reaction containing RNA or DNA is mixed with Binding Solution and the nucleic acids are bound to BSI's column (BIND). Under these conditions only the nucleic acids will bind to BSI's resin while most of the contaminating proteins and nucleotides are removed in the flowthrough. The bound nucleic acid is then washed to remove any remaining impurities (WASH). Lastly, the purified total RNA or DNA is eluted into 50 µL of the provided Elution Buffer or water (ELUTE). Please see procedure flowchart to the right.


FEATURES AND BENEFITS

* Versatile performance - Both DNA and RNA can be cleaned and concentrated using the same kit. Clean-up nucleic acids from a variety of isolation methods and upstream enzymatic reactions.
* Fast and easy processing - Rapid spin-column format allows for the processing of 10 samples in 20 minutes.
* Suitable for all sizes of RNA - Purify all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA).
* Suitable for all sizes of DNA - Purify all sizes of DNA, from small PCR products, native or linearized plasmids, to genomic DNA. Using an alternative protocol, even oligonucleotides and smaller DNA fragments can be purified.
* Optional endotoxin removal - Optional protocol is provided for the rapid removal of endotoxins from previously purified RNA or DNA.
* No phenol:chlorofom extractions - BSI’s CleanAll kit purifies RNA without the use of harmful chemicals such as phenol or chloroform

APPLICATIONS AND DATA

The kit can be used to clean up RNA and DNA from upstream applications such as:

* Digestions
* Ligations
* PCR reactions
* Labeling reactions
* DNA modification reactions
* Staining

Figure 1. High Recoveries and Efficient Endotoxin Removal. High recoveries of DNA can be seen when using the CleanAll kit, and these high recoveries are not compromised even during endotoxin removal. In Panel A, 1 µg of a PCR product was cleaned using the CleanAll RNA/DNA Clean-Up and Concentration Kit as well as BSI's PCR Purification Kit. More than 0.9 µg of the DNA was recovered from both kits with a comparable percent recovery of over 90%. In Panel B, 10 µg of plasmid DNA preparation was cleaned from endotoxin by using the CleanAll RNA/DNA Kit. Endotoxin levels of the plasmid DNA sample were reduced from 0.75 EU/µg plasmid DNA to less 0.05 EU/µg plasmid DNA. Approximately 95% of the endotoxin units in the DNA sample were removed using the kit, therefore the kit allows for high recoveries coupled with exceptional cleanup.

Figure 2: Clean-Up of RNA with High Recovery. BSI’s CleanAll RNA/DNA Clean-Up and Concentration Kit can be used to clean up various enzymatic reactions including DNase treatment. In Figure 2 above, Lane 1 is the RNA input, while lanes 2-5 contain the RNA that has been cleaned using BSI’s CleanAll Kit. It can be seen that in all cases the recovery is high, and the purified RNA is intact and of a high quality.

CleanAll RNA/DNA Clean-Up and Concentration Kit Components

1. Binding Solution
2. Wash Solution
3. Elution Buffer
4. Micro Spin Columns
5. Collection Tubes
6. Elution tubes
7. Product Insert

Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.

Protein Purification Kit

4-in-1 RNA/miRNA/DNA/Protein Purification Kit

Catalog# : NB-24200
Price/Unit : $295.00
Unit : 20 preps
Purify RNA, microRNA, total proteins and genomic DNA from the same sample

* Purify RNA, microRNA, proteins and DNA from a single sample
* No phenol extraction step
* Fast and easy processing
* Ideal for precious, difficult to obtain or small samples
* Flexible protocol allows for total RNA purificaiton (including microRNA) or separate enrichment of microRNAs

Bio-Synthesis All-in-One Purification Kit provides a rapid method for the isolation and purification of total RNA including microRNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants. In addition, a microRNA Enrichment Column is provided for the optional seperate purification of small RNA (<200>200 nt) and the small/microRNA (<200>FEATURES AND BENEFITS

* No phenol - Isolate RNA, DNA and proteins without the use of phenol or chloroform
* Complete column purification Isolate total or micro-enriched RNA- Two different protocols are provided to isolate either total RNA, including all sizes of RNA from large rRNA to microRNA, or separately isolate microRNA species (<200>200 nt)
* Reduce variability - RNA, DNA and proteins are isolated from a single sample with no splitting of the lysate, thus reducing inconsistent results and variability.
* Isolate from small samples - Sequential isolation of RNA, DNA and protein from a single sample. Ideal for precious, difficult to obtain or small samples.
* Rapid procedure - Isolate total RNA, genomic DNA and total proteins OR microRNA, large RNA, genomic DNA and proteins from a single sample in <>Figure 1. Sequential Isolation of microRNA, Large RNA, Genomic DNA and Proteins from 1 x 106 HeLa Cells. Panels A and B show the separate isolation of large RNA and microRNA from 2 different samples of HeLa cells. Panel A is a 1X MOPS 1% agarose gel and Panel B is a 10% urea-PAGE gel. In both gels, Lane M is Bio-Synthesis's 1Kb RNA Ladder, Lanes 1 and 2 contain 3 µL out of the 50 µL elutions of the large RNA fraction, and Lanes 3 and 4 contain 3 µL out of the 50 µL elutions of the microRNA fractions. Panel C is a 1% agarose gel showing the gDNA isolated from the same 2 HeLa cell samples. Lane M is Bio-Synthesis's UltraRanger DNA Ladder and Lanes 1 and 2 contain 10 µL of each of the 100 µL elutions. Panel D is a 12% SDS-PAGE gel that contains the proteins that were isolated from the 2 HeLa cell samples. Lane M is a protein ladder and Lanes 1 and 2 contain 10 µL of the 100 µL elution of proteins that had been column purified. The large RNA, microRNA, gDNA and proteins are all intact and of the highest integrity and quality. Furthermore, the kit allows for the successful separate isolation of microRNA (<200>200nt).

Figure 2. Sequential Isolation of microRNA, Large RNA, Genomic DNA and Proteins from 5 x 106 Yeast Cells. Panels A shows the separate isolation of large RNA and microRNA from a plasmid-containing yeast culture. Panel A depicts the resolution of 1 µL of the 50 µL large RNA (Top) and small RNA (Bottom) fractions on an Agilent RNA Nano 6000 Chip. Panel B is a 1% agarose gel showing 10 µL of 100 µL of the gDNA and plasmid DNA isolated from the same yeast samples with Bio-Synthesis's UltraRanger DNA Ladder as molecular weight marker. Panel C is a 12% SDS-PAGE gel that contains 10 µL of 100 µL of the proteins that were isolated from the yeast samples. The large RNA, microRNA, gDNA and proteins are all intact and of the highest integrity and quality. Furthermore, the kit allows for the successful separate isolation of microRNA (<200>200nt) with the complete elimination of large rRNA in the small RNA fraction.
Figure 3. Isolate High Quality RNA. RNA was isolated from 0.8 million HeLa cells using Bio-Synthesis's All-in-One Purification Kit and a leading market competitor for multiple analyte isolations. Half a microgram of the purified RNA (50 µL elution volume) was used as the template in a 20 µL reverse transcription reaction using oligo dT primers. One microliter of the RT reaction was used in a 20 µL qPCR reaction using the human GAPDH primers, and the results are shown in the PCR baseline graph. The blue lines correspond to the PCR results when RNA isolated using Bio-Synthesis's kit was used as the template, while the green lines correspond to the results when RNA isolated using the competitors kit was used as the template. From the graph it can be seen that Bio-Synthesis's kit isolated RNA with a greater sensitivity, and that more RNA was isolated from the same input as evidenced by the lower Ct values for the Bio-Synthesis-isolated RNA.

All-in-One Purification Kit (Total RNA, microRNA, total proteins and genomic DNA) Components

1. Lysis Solution
2. RNA Wash Solution
3. RNA Elution Solution
4. gDNA Wash Solution
5. gDNA Elution Buffer
6. Protein Wash Buffer
7. Protein pH Binding Buffer
8. Protein Elution Buffer
9. Protein Neutralizer
10. Protein Loading Dye
11. All-in-One Spin Columns
12. microRNA Enrichment Column
13. Collection Tubes
14. Elution tubes
15. Product Insert

Long-Term Storage : The Protein Loading Dye should be stored at -20°C upon arrival. All other solutions should be kept tightly sealed and stored at room temperature. These reagents should remain stable for at least 1 year in their unopened containers
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.

Custom DNA Peptide-Based Assay Development

Custom DNA Peptide-Based Assay Development

As researchers approach the developing field of genomics with custom DNA/peptide based assays, the need for microarray services or productions that maximize flexibility and precision, without compromising speed and cost, has become critical. In order to meet the demands for custom microarraying, BSI offers a complete suite of microarray services, which includes:

• Experimental design consultation, provide expert support with DNA/peptide sequence selection, microarray chip layout design, and quality control.
• Synthesis chemistry of custom DNA, RNA, peptide, BNA or hybrid oligos with backbone modifications such as phosphorothioate for antisense study, or nucleotide modifications
• Flexible chip fabrication, which allow fine-tune client's probe sets for high-density custom microarrays.
• Target sample labeling and hybridization
• Other related services DNA/RNA Extraction, RNA quality check, Real-Time PCR, Western blot.
• BioInformatic data analysis

For complete details or to get a specific quotation, please contact a Technical Support Representative by submitting an Online Oligo Project Requisition Form. Clients must provide information such as gene ID list, or send u s oligo/peptide/protein samples. A Customer Service representative will contact you as soon as possible to finalize your array design and price quotation. Customized arrays will be shipped in 2-3 weeks upon finalization of the order.

To download more specific instructions and an appropriate order form, please click here.

Quality Control

Replicated spots and slides are used to monitor technical variations. In addition, pre-scanning with fast dye staining is conducted to make sure the spot size and morphology meet expectations. Sometimes a test hybridization is conducted, if needed. Using similar high-precision printing technology, the peptides or proteins can be spotted on membrane-coated glass slides. Using different detection technologies, recombinant proteins/antibodies, cell lysate or serum samples can be quantitatively assayed. This has wide applications in protein-protein interaction studies, including antigen-antibody recognitions, kinase-substrate bindings, pathway analysis, biomarker screenings, and much more.

Services highlight

• DNA/Peptide selection and modification
• sample to data full service

FAIM Antibody

FAIM Antibody

Catalog# : 2309

The susceptibility of primary splenic B cells to Fas-mediated apoptosis is regulated in a receptor-specific fashion. Terminal effectors of B cell Fas-resistance include the known anti-apoptotic proteins Bcl-xL, FLIP, and a recently identified protein termed FAIM. This molecule is broadly expressed in various tissues and exists in at least three isoforms. It is thought that resistance to Fas killing via increased expression of FAIM protects foreign antigen-specific B cells during interactions with FasL-bearing T cells whereas autoreactive B cells are deleted via Fas-dependent cytotoxicity. More recent results have indicated that FAIM interacts with both Trk and p75 neurotrophin receptor and may play a role in promoting neurite outgrowth in different neuronal systems by a mechanism involving the activation of NF-kappaB and the Ras-ERK pathway.

Additional Names : FAIM, Fas apoptotic inhibitory molecule
Description
Left: Western blot analysis of FAIM in human spleen tissue lysate with FAIM antibody at (A) 5 and (B) 10 µg/ml.





Source : FAIM antibody was raised against a 14 amino acid peptide from near the carboxy terminus of human FAIM .

Purification : Affinity chromatography purified via peptide column

Clonality and Clone : This is a polyclonal antibody.

Host : FAIM antibody was raised in rabbit. Please use anti-rabbit secondary antibodies.

Application : FAIM antibody can be used for detection of FAIM by Western blot at 5 – 10 µg/ml.

Tested Application(s) : E, WB

Buffer : Antibody is supplied in PBS containing 0.02% sodium azide.

Blocking Peptide : Cat.No. 2309P - FAIM Peptide

Long-Term Storage : FAIM antibody can be stored at 4ºC, stable for one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

Positive Control
1. Cat. No. 1306 - Human Spleen Tissue Lysate

Species Reactivity :H, M

GI Number : 8922536

Accession Number : NP_060617

Short Description : an apoptosis inhibitor

References
1. Rothstein TL. Inducible resistance to Fas-mediated apoptosis in B cells. Cell Res. 2000; 10:245-66.
2. Schneider TJ, Fischer GM, Donohoe TJ, et al. A novel gene coding for a Fas apoptosis inhibitory molecule (FAIM) isolated from inducibly Fas-resistant B lymphocytes. J. Exp. Med. 1999; 189:949-55.
3. Sole C, Dolcet X, Segura MF, et al. The death receptor antagonist FAIM promotes neurite outgrowth by a mechanism that depends on ERK and NF-kappa B signaling. J. Cell Biol. 2004; 167:479-92.

FAF1 Antibody

FAF1 Antibody

Catalog# : 3993

Fas-associated protein 1 (FAF1) was initially identified as a Fas-binding pro-apoptotic protein that is component of the death-inducing signaling complex in Fas-mediated apoptosis (1). FAF1 can also induce apoptosis in the absence of extrinsic death signals when overexpressed although it does not contain typical death motifs such as the death domain, death effector domain, and caspase recruitment domain (1,2). Overexpression of FAF1 also decreases the basal level of NF-kappaB activity in transfected 293 cells, inhibits NF-kappaB activity induced by TNF-alpha, IL-1beta and lipopolysaccharide, and prevents NF-kappaB translocation to the nucleus (3), suggesting that another role of FAF1 is to negatively regulate the activity of NF-kappaB. FAF1 can also interact with the inflammatory signaling PYRIN-containing Apaf-1-like proteins (PYPAFs, also called NALPs) such as PYPAF1, PYPAF2 (NALP2), and PYPAF7, suggesting FAF1 may also be involved in the inflammation pathway (4). Multiple differentially spliced isoforms of FAF1 are known to exist.

Additional Names : FAF1 (CT), Fas-associated protein 1, hFAF1
Description
Left: Western blot analysis of FAF1 in Jurkat cell lysate with FAF1 antibody at (A) 1 and (B) 2 µg/ml.

Below: Immunohistochemistry of FAF1 in rat spleen tissue with FAF1 antibody at 2.5 µg/ml.

Other Product Images

Source : FAF1 antibody was raised against a 19 amino acid peptide from near the carboxy terminus of human FAF1.

Purification : Affinity chromatography purified via peptide column

Clonality and Clone : This is a polyclonal antibody.

Host : FAF1 antibody was raised in rabbit. Please use anti-rabbit secondary antibodies.

Application : FAF1 antibody can be used for detection of FAF1 by Western blot at 1 – 2 µg/ml.

Tested Application(s) : E, WB, IHC

Buffer : Antibody is supplied in PBS containing 0.02% sodium azide.

Blocking Peptide : Cat.No. 3993P - FAF1 Peptide

Long-Term Storage : FAF1 antibody can be stored at 4ºC, stable for one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

Positive Control
1. Cat. No. 1205 - Jurkat Cell Lysate
2. Cat. No. 1466 - Rat Spleen Tissue Lysate

Species Reactivity :H, M, R

GI Number : 5901948

Accession Number : NP_008982

Short Description : (CT) Fas-associated protein 1

References
1. Chu K, Niu X, and Williams LT. A Fas-associated protein factor, FAF1, potentiates Fas-mediated apoptosis. Proc. Natl. Acad. Sci. USA 1995; 92:11894-8.
2. Ryu SW and Kim E. Apoptosis induced by human Fas-associated factor 1, hFAF1, requires its ubiquitin homologous domain, but not the Fas-binding domain. Biochem. Biophys. Res. Commun. 2001; 286:1027-32.
3. Park M-Y, Jang HD, Lee SY, et al. Fas-associated Factor-1 inhibits Nuclear Factor-kappaB (NF-kappaB) activity by interfering with nuclear translocation of the RelA (p65) subunit of NF-kappaB. J. Biol. Chem. 2004; 279:2544-9.
4. Kinoshita T, Kondoh C, Hasegawa M, et al. Fas-associated factor 1 is a negative regulator of PYRIN-containing Apaf-1-like protein 1. Int. Immunol. 2006;18:1701-6.

F1A alpha Antibody

F1A alpha Antibody

Catalog# : 2279

Fas and tumor necrosis factor receptor 1 (TNFR1) are two prototype members in the death receptor family. A novel protein that associates with the intracellular domains of Fas and TNFR1 was recently identified and designated F1Aalpha and FEM1b (1,2). F1Aalpha/FEM1beta is the homologue of C. elegans sex determining protein FEM-1 (1,2). FEM-1/F1Aalpha is cleaved by CED-3 and caspase (3). FEM-1/F1Aalpha associates with CED-4 and its mammalian homologue Apaf-1 (3). Overexpression of F1Aalpha induces apoptosis. F1Aalpha is therefore a novel member of the death receptor associated protein that mediates apoptosis. F1Aalpha is expressed in a variety of human and mouse tissues (1,2)

Additional Names : F1A alpha (CT), F1A alpha
Description
Left: Western blot analysis of F1A alpha in mouse (A) and rat (B) liver tissue lysates with F1A alpha antibody at 1 µg/ml.

Below: Immunohistochemistry of F1Aalpha in mouse liver tissue with F1Aalpha antibody at 5 µg/ml.

Other Product Images

Source : F1A alpha antibody was raised against a peptide corresponding to amino acids 609 to 622 of human F1A alpha. This sequence is identical to the corresponding sequence of mouse FEM1b .

Purification : Affinity chromatography purified via peptide column

Clonality and Clone : This is a polyclonal antibody.

Host : F1A alpha antibody was raised in rabbit. Please use anti-rabbit secondary antibodies.

Immunogen : Human F1A alpha Peptide (Cat. No. 2279P)

Application : F1Ab antibody can be used for detection of F1Ab by Western blot at 0.5 to 1 µg/ml.An approximately 70 kDa band can be detected. It is human, mouse, and rat reactive.

Tested Application(s) : E, WB, IHC

Buffer : Antibody is supplied in PBS containing 0.02% sodium azide.

Blocking Peptide : Cat. No. 2279P - F1A alpha Peptide

Long-Term Storage : F1A alpha antibody can be stored at 4ºC, stable for one year. As with all antibodies care should be taken to avoid repeated freeze thaw cycles. Antibodies should not be exposed to prolonged high temperatures.

Positive Control
1. Cat. No. 1404 - Mouse Liver Tissue Lysate
2. Cat. No. 1464 - Rat Liver Tissue Lysate

Species Reactivity :H, M, R

GI Number : 6175869

Accession Number : AAF05314

Short Description : (CT) A novel death receptor-binding protein

References
1. Chan SL, Tan KO, Zhang L, Yee KS , Ronca F, Chan MY, Yu VC. F1Aalpha, a death receptor-binding protein homologous to the Caenorhabditis elegans sex-determining protein, FEM-1, is a caspase substrate that mediates apoptosis. J Biol Chem. 1999;274(45):32461-8.
2. Ventura-Holman T, Maher JF. Sequence, organization, and expression of the human FEM1B gene. Biochem Biophys Res Commun 2000;267(1):317-20
3. Chan SL, Yee KS , Tan KM, Yu VC. The Caenorhabditis elegans sex determination protein FEM-1 is a CED-3 substrate that associates with CED-4 and mediates apoptosis in mammalian cells. J Biol Chem. 2000;275(24):17925-8. (WD0102)

EndoG Monoclonal Antibody

EndoG Monoclonal Antibody

Catalog# : PM-4579

The fragmentation of nuclear DNA is a hallmark of apoptotic cell death. The activities of caspase and nuclease are involved in the DNA fragmentation. Caspase-activated deoxyribonuclease (CAD), also termed DNA fragmentation factor (DFF40), is one such nuclease, and is capable of inducing DNA fragmentation and chromatin condensation after cleavage by caspase-3 of its inhibitor ICAD/DFF45. Caspase and CAD independent DNA fragmentation also exists. Recent studies demonstrated that another nuclease, endonuclease G (EndoG), is specifically activated by apoptotic stimuli and is able to induce nucleosomal fragmentation of DNA independently of caspase and DFF/CAD. EndoG is a mitochondrion-specific nuclease that translocates to the nucleus and cleaves chromatin DNA during apoptosis. The homologue of mammalian EndoG is the first mitochondrial protein identified to be involved in apoptosis in C. elegans. EndoG also cleaves DNA in vitro.

Additional Names : EndoG (7G1G10): Endonuclease G

Description
Left: Western blot analysis of EndoG expression in HepG2 cell lysate with EndoG antibody at (A) 5 and (B) 10 µg/ml. a





Source : Mouse monoclonal EndoG antibody was raised against a recombinant protein corresponding to amino acids 51 – 140 of human EndoG.

Purification : Immunoaffinity chromotography purified IgG

Clonality and Clone : This is a monoclonal antibody. (Clone 7G1G10)

Host : EndoG monoclonal antibody was raised in mouse. Please use anti-rabbit secondary antibodies.

Immunogen : Recombinant protein corresponding to amino acids 51 – 140 of human EndoG.

Application : EndoG monoclonal antibody can be used for detection of EndoG by Western blot at 5 – 10 µg/ml.

Tested Application(s) : E, WB

Buffer : Antibody is supplied in PBS containing 0.02% sodium azide.

Blocking Peptide :
Long-Term Storage : EndoG monoclonal antibody can be stored at 4ºC, stable for one year.

Positive Control
1. Cat. No. 1211 - HepG2 Cell Lysate
2. Cat. No. 95-103 - EndoG Recombinant Protein

Species Reactivity :H, R

Short Description : Endonclease G

References
1. Li LY, Luo X, Wang X. Endonuclease G is an apoptotic DNase when released from mitochondria. Nature 2001; 412:95-9.
2. Parrish J, Li L, Klotz K, et al. Mitochondrial endonuclease G is important for apoptosis in C. elegans. Nature 2001; 412:90-4
3. Hengartner MO. Apoptosis. DNA destroyers. Nature 2001; 412:27, 29.
4. Widlak P, Li LY, Wang X, et al. Action of recombinant human apoptotic endonuclease G on naked DNA and chromatin substrates: cooperation with exonuclease and DNase I. J. Biol. Chem. 2001; 276:48404-9.

Use of Bioconjugates in Apoptosis

Use of Bioconjugates in Apoptosis

Bioconjugation Service: In some cases the carrier moiety can be a synthetic polymer such as poly-L-glutamic acid to which the drug paclitaxel has been conjugated. The actual mechanism of action is still being elucidated. Another carrier that has been used successfully are the cyclodextrins to which small pro-apoptotic agents can be linked. For example For instance a conjugate of anti-CD33 antibody and the amphipathic peptide KLA target efficiently CD33-positive myeloid leukemia cell lines to cause their apoptotic death induced by the D-(KLAKLAK)2 proapoptotic peptide.

Bioconjugates: The study of apotosis, the programmed death of cells, has direct applications to cancer a disease where tumor cells have developed mechanisms to avoid it and multiply without control. The fact that apoptosis is usually mediated by some cell receptors makes conjugates a valuable tool in elucidating apoptosis’ mechanisms as well drug development.

Bioconjugations: Cells over-expressing erbB2 and resistant to apoptosis can be killed more efficiently by a conjugate composed of an erbB2-binding heptapeptide conjugated to the proapoptotic a-tocopheryl succinate (a-TOS) rather than the unconjugated a-TOS. Use of the conjugate resulted in breast carcinomas in a breast cancer prone transgenic mouse strain. Recently it has been shown that conjugates of cytochrome c and oligoarginine linked by a thioether resulted in an increased entry into the cell, compared to cyt. c alone, and increase in apoptotic activity. In contrast a conjugate linked by a disulfide bond although entered into the cell did not enhance the apoptotic activity.

Protein Purification Kit

4-in-1 RNA/miRNA/DNA/Protein Purification Kit

Catalog# : NB-24200
Price/Unit : $295.00
Unit : 20 preps
Purify RNA, microRNA, total proteins and genomic DNA from the same sample

* Purify RNA, microRNA, proteins and DNA from a single sample
* No phenol extraction step
* Fast and easy processing
* Ideal for precious, difficult to obtain or small samples
* Flexible protocol allows for total RNA purificaiton (including microRNA) or separate enrichment of microRNAs

Bio-Synthesis's All-in-One Purification Kit provides a rapid method for the isolation and purification of total RNA including microRNA, genomic DNA and proteins sequentially from a single sample of cultured animal cells, small tissue samples, blood, bacteria, yeast, fungi or plants. In addition, a microRNA Enrichment Column is provided for the optional seperate purification of small RNA (<200 nt), allowing for 4-in-1 purification (large RNA, genomic DNA, microRNA and proteins). This kit is an ideal all-in-one solution for researchers studying systems biology, including those who are interested in the interactions of multiple disciplines including RNA interference, genomics, epigenomics, transcriptomics and proteomics. Bio-Synthesis's All-in-One Purification Kit is especially useful for researchers who are isolating macromolecules from precious, difficult to obtain or small samples such as needle biopsies and single foci from cell cultures, as well as for mutant analysis, RNA interference studies and pathogen detection. Furthermore, analysis will be more reliable since the RNA, DNA and proteins are derived from the same sample without any fractionation, thereby eliminating inconsistent results. The purification procedure is very rapid, allowing for the isolation of large RNA, genomic DNA, microRNA and proteins from a single sample in less than 40 minutes. The isolated macromolecules are of the highest purity and can be used in a number of different downstream applications.

Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. Briefly, the process involves first lysing the cells or tissue of interest with the provided Lysis Solution. Ethanol is then added to the lysate, and the solution is loaded onto a spin-column. Under these conditions only the RNA and DNA will bind to Bio-Synthesis's resin, while the proteins are removed in the flowthrough (BIND 1). An additional protocol utilizing the microRNA Enrichment Column is provided if the user wishes to further separate the large (>200 nt) and the small/microRNA (<200 nt). Next, the bound RNA is washed with the provided RNA Wash Solution to remove impurities, and the purified RNA is eluted with the RNA Elution Solution (WASH ELUTE 1A). The remaining bound genomic DNA is then washed with the provided gDNA Wash Buffer to remove any remaining trace levels of RNA, and the gDNA is then eluted with the gDNA Elution Buffer (WASH ELUTE 1B). The kit purifies all sizes of RNA, from large mRNA and ribosomal RNA down to microRNA (miRNA) and small interfering RNA (siRNA).

The proteins that are present from the initial flowthrough can now be loaded directly onto an SDS-PAGE gel for visual analysis. Alternatively, the protein samples can be further purified using the spin columns provided with the kit. After the RNA and gDNA has been eluted from the column, the flowthrough is then pH adjusted and loaded back onto the column in order to bind the proteins that are present (BIND 2). The bound proteins are washed with the provided wash buffer (WASH 2), and are then eluted such that they can be used in downstream applications (ELUTE 2).

The purified RNA, DNA and proteins are of the highest quality and can be used in a number of downstream applications.FEATURES AND BENEFITS

* No phenol - Isolate RNA, DNA and proteins without the use of phenol or chloroform
* Complete column purification Isolate total or micro-enriched RNA- Two different protocols are provided to isolate either total RNA, including all sizes of RNA from large rRNA to microRNA, or separately isolate microRNA species (<200>200 nt)
* Reduce variability - RNA, DNA and proteins are isolated from a single sample with no splitting of the lysate, thus reducing inconsistent results and variability.
* Isolate from small samples - Sequential isolation of RNA, DNA and protein from a single sample. Ideal for precious, difficult to obtain or small samples.
* Rapid procedure - Isolate total RNA, genomic DNA and total proteins OR microRNA, large RNA, genomic DNA and proteins from a single sample in < 40 minutes.
* Isolate a diversity of RNA species - All sizes of RNA are isolated, from large mRNA down to microRNA, either separately or combined in a total RNA fraction.
* Process a wide range of sample types - Isolate total RNA, genomic DNA and proteins from cultured animal cells , tissue, blood, bacteria, yeast, fungi and plants.

APPLICATIONS

RNA

* Bioanalyzer
* Quantitative, real-time RT-PCR for both large mRNA and small RNA including miRNA
* RT-PCR for both large mRNA and small RNA including miRNA
* Northern blotting
* RNase protection
* Primer extension
* Expression array assays
* Next Generation Sequencing
* microRNA cloning
* PCR-based virus detection
* PCR-based viable bacteria detection

Genomic DNA

* PCR and Quantitative PCR
* Sequencing
* DNA methylation studies
* Southern blotting
* Microarrays
* SNP analysis
* PCR-based virus detection/identification
* PCR-based bacteria detection/identification

Protein

* SDS-PAGE analysis
* Western blots
* Mass spectrometry Figure 1. Sequential Isolation of microRNA, Large RNA, Genomic DNA and Proteins from 1 x 106 HeLa Cells. Panels A and B show the separate isolation of large RNA and microRNA from 2 different samples of HeLa cells. Panel A is a 1X MOPS 1% agarose gel and Panel B is a 10% urea-PAGE gel. In both gels, Lane M is Bio-Synthesis's 1Kb RNA Ladder, Lanes 1 and 2 contain 3 µL out of the 50 µL elutions of the large RNA fraction, and Lanes 3 and 4 contain 3 µL out of the 50 µL elutions of the microRNA fractions. Panel C is a 1% agarose gel showing the gDNA isolated from the same 2 HeLa cell samples. Lane M is Bio-Synthesis's UltraRanger DNA Ladder and Lanes 1 and 2 contain 10 µL of each of the 100 µL elutions. Panel D is a 12% SDS-PAGE gel that contains the proteins that were isolated from the 2 HeLa cell samples. Lane M is a protein ladder and Lanes 1 and 2 contain 10 µL of the 100 µL elution of proteins that had been column purified. The large RNA, microRNA, gDNA and proteins are all intact and of the highest integrity and quality. Furthermore, the kit allows for the successful separate isolation of microRNA (<200>200nt).

Inclusion Body Solubilization Reagent

Inclusion Body Solubilization Reagent

Catalog# : NB-18700
Price/Unit : $71.80
Unit : 25 mL

For the efficient solubilization of inclusion body proteins

* Fast dissolving of inclusion body aggregates
* Allows for downstream applications of inclusion body proteins

Bio-Synthesis's Inclusion Body Solubilization Reagent dissolves inclusion body aggregates resulting from the expression of recombinant proteins in bacteria. This reagent has demonstrated exceptional ablility to solubilize inclusion bodies. Once dissolved, proteins can be analyzed by SDS-PAGE, quantified or further purified for refolding.

Inclusion bodies can be released from bacterial cells using Bio-Synthesis's Cell Lysis Reagent (available separately). Bio-Synthesis's Inclusion Body Protein Isolation Kits can also be used as complete kits for cell disruption, inclusion body solubilisation and purification using spin column chromatography

APPLICATIONS

Solubilized proteins can be used in a number of downstream applications including:

* SDS-PAGE analysis
* Western blot
* ELISA
* Refolding experiments
* Mass spectrometry
* Further purification and scale up

Figure 1: Efficient Isolation of Inclusion Body Proteins Using Bio-Synthesis's Cell Lysis Reagent and Inclusion Body Solubilization Reagent. Inclusion Bodies were extracted and solubilized following gene expression. Recovered proteins were analyzed on 12% SDS-PAGE and stained with Coomassie Brilliant Blue R-250. Numbers represent kDa sizes of protein bands.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.

Cell Lysis Reagent

Cell Lysis Reagent

Catalog# : NB-18800
Price/Unit : $110.00
Unit : 100 mL

For the efficient lysis of bacterial cells and the extraction of inclusion body proteins

* Proprietary solution of detergents, protease-inhibitors and buffer
* Efficient lysis of bacterial cells for extraction of inclusion body proteins

Bio-Synthesis's Cell Lysis Reagent is designed for the gentle and efficient lysis of bacterial cells in order to assist in the extraction of inclusion body proteins. Cell lysis is accomplished through non-ionic detergent chemical disruption in conjunction with mechanical disruption. The use of a needle and syringe during the procedure helps to reduce viscosity and facilitate purification, producing proteins that are often greater than 95% pure.

Cell Lysis Reagent does not solubilize inclusion bodies. Inclusion Bodies can be solubilized using Bio-Synthesis's Inclusion Body Solubilization Reagent (available separately). Bio-Synthesis's Inclusion Body Protein Isolation Kits can also be used as complete kits for cell disruption, inclusion body solubilisation and purification using spin column chromatography.

APPLICATIONS
Proteins purified using Bio-Synthesis's Cell Lysis Reagent can be used in a number of downstream applications including:

* SDS-PAGE analysis
* Western blot
* ELISA
* Refolding experiments
* Mass spectrometry
* Further purification and scale up

Figure 1: Efficient Isolation of Inclusion Body Proteins Using Bio-Synthesis's Cell Lysis Reagent and Inclusion Body Solubilization Reagent. Inclusion Bodies were extracted and solubilized following gene expression. Recovered proteins were analyzed on 12% SDS-PAGE and stained with Coomassie Brilliant Blue R-250. Numbers represent kDa sizes of protein bands

Customer-supplied Reagents and Equipment

* Centrifuge
* Micropipettors
* Needles (20 guage for small scale protocol and 18 guage for large scale protocol)
* Syringes (1 mL for small scale protocol, 10 mL for large scale protocol)
* Sterile deionized or Milli-Q ® water

Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.

ProteoSpin Total Protein Detergent Clean-Up Micro Kit

ProteoSpin Total Protein Detergent Clean-Up Micro Kit

Catalog# : NB-23300
Purity : Purity > 95% by HPLC
Solubility : Distilled water for a solution up to 2 mg/ml, otherwise we recommend using DMSO
Format : Each vial contains lyophilized solid packaged and supplied as a trifluoroacetate salt
Price/Unit : $230.00
Unit : 20 preps
For the rapid concentration and clean-up of protein samples

* Concentrate protein samples and remove detergents at the same time
* Rapid and convenient spin-column protocol
* Effectively remove a wide range of detergents including SDS, Triton® X-100, CHAPS, NP-40, and Tween 20

The ProteoSpinTM Total Protein Detergent Clean-Up Micro Kit concentrates proteins from lysates and removes detergents, denturants, and other ingredients commonly employed in the lysis of cells and tissues, while concentrating the proteins at the same time. The kit employs a novel spin column technology that offers convenience, speed, and ease-of-use. The kit can remove trace amounts of detergents from total protein samples while maintaining high protein recovery. All types of detergents including ionic, non-ionic and zwitterionic detergents can be effeciently removed in a very short time eliminating the need for for labourious procedures such as dialysis. The kit is also designed to remove detergents in protein solutions either in their free form or bound form, as when complexed with the protein. The ProteoSpinTM Total Protein Detergent Clean-Up Micro Kit removes a variety of detergents and denturants at the same time, unlike other commercially avialable kits that can only handle one detergent and often don’t clean up denaturants. This is particularly important since lysis reagents often employ a combination of detergent(s) and denturants for efficient cell lysis. No further cleaning of the protein sample is required, and it can be used directly in a variety of downstream applications including mass spectrometry, SDS-PAGE, isoelectric focusing, and NMR spectroscopy.

The kit employs a prorietary matrix to remove detergents from protein samples. Breifly, pH Binding Buffer is added to the protein sample and the sample is applied to Bio-Synthesis's column (BIND). Under these conditions the proteins will bind to the column while detergents and other contaminants are removed in the flowthrough. The bound proteins are then washed to remove any remaining impurities (WASH). Lastly, the clean proteins are eluted into a small volume of the provided Elution Buffer or into other optional elution buffers (user-provided) (ELUTE). Please see the procedure flowchart to the right.
FEATURES AND BENEFITS

* Reomve a variety of detergents - Detergents including SDS, Triton® X-100, CHAPS, NP-40, Triton® X-114 and Tween 20 can be removed using the kit.
* Effective detergent removal - Detergent removal of greater than 95% for many protein samples, allowing for detergent-sensitive tryptic digestion. Can also remove detergents that are complexed with proteins.
* Complete kit - The ProteoSpinTM Total Protein Detergent Clean-Up Micro Kit contains all the solutions and columns for the processing of 20 total protein samples.
* Fast and easy processing - Preparation time for 12 samples is only 20 minutes.
* Eluted proteins can be used directly in downstream applications - No need for additional sample preparation procedures for downstream applications. Concentrate protein sample and remove detergents at the same time.
* No detergent carryover - Each sample is processed individually using its own spin column, resulting in no carryover or column bleed from sample to sample.
* Ready-to-use columns - No purging, swelling of resin, or lengthy activation steps required.


APPLICATIONS
Concentrated, detergent-free proteins are suitable for a number of downstream applications such as:

* 2D SDS-PAGE
* Whole protein mass spectrometry
* Isoelectric focusing
* X-ray crystallography
* NMR spectroscopy
* Protein microarrays
Detergent Clean-Up Micro Kit Contents

1.Binding Buffer
2.Wash Buffer I
3.Wash Buffer II
4.Elution Buffer
5.Neutralizer
6.Micro Spin Columns (20)
7.Collection Tubes (20)
8.Elution Tubes (20)
9.Product Insert

Long-Term Storage : Unopened solutions should be stored at room temperature. Once opened, the solutions should be stored at 4°C when not in use.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.

Specialty Peptide Synthesis

Short Peptide Molecule Synthesis

BSI offers short peptide synthesis that are designed from 2-3 amino acid for potentially large quantities. Different synthesis strategies and chemistry will applied with the complexity of the sequence (for example hydrophobic, short peptides) or quantity yield.

For more information about our short and large quantity peptide synthesis, Please call 800-227-0627 or write to us at biosyn@biosyn.com

Service Features:

All custom
short peptide synthesis includes:

• HPLC for purity
• MALDI-TOF mass spectrometry for identification and characterization
• Certificate of Analysis.
  

Additional services: amino acid analysis and sequencing

Please call 800-227-0627 or write to us at biosyn@biosyn.com for details

For other peptide synthesis services, click here!

EndoG Monoclonal Antibody

EndoG Monoclonal Antibody

Catalog# : PM-4581

The fragmentation of nuclear DNA is a hallmark of apoptotic cell death. The activities of caspase and nuclease are involved in the DNA fragmentation. Caspase-activated deoxyribonuclease (CAD), also termed DNA fragmentation factor (DFF40), is one such nuclease, and is capable of inducing DNA fragmentation and chromatin condensation after cleavage by caspase-3 of its inhibitor ICAD/DFF45. Caspase and CAD independent DNA fragmentation also exists. Recent studies demonstrated that another nuclease, endonuclease G (EndoG), is specifically activated by apoptotic stimuli and is able to induce nucleosomal fragmentation of DNA independently of caspase and DFF/CAD. EndoG is a mitochondrion-specific nuclease that translocates to the nucleus and cleaves chromatin DNA during apoptosis. The homologue of mammalian EndoG is the first mitochondrial protein identified to be involved in apoptosis in C. elegans. EndoG also cleaves DNA in vitro.

Additional Names : EndoG (7F2D7): Endonuclease G
Source : Mouse monoclonal EndoG antibody was raised against a recombinant protein corresponding to amino acids 51 – 140 of human EndoG.






Source : Mouse monoclonal EndoG antibody was raised against a recombinant protein corresponding to amino acids 51 – 140 of human EndoG.

Purification : Immunoaffinity chromotography purified IgG

Clonality and Clone : This is a monoclonal antibody. (Clone 7F2D7)

Host : EndoG monoclonal antibody was raised in mouse. Please use anti-rabbit secondary antibodies.

Immunogen : Recombinant protein corresponding to amino acids 51 – 140 of human EndoG.

Application : EndoG monoclonal antibody can be used for detection of EndoG by Western blot at 5 – 10 µg/ml.

Tested Application(s) : E, WB

Buffer : Antibody is supplied in PBS containing 0.02% sodium azide.

Long-Term Storage : EndoG monoclonal antibody can be stored at 4ºC, stable for one year.

Positive Control
1.Cat. No. 1211 - HepG2 Cell Lysate
2.Cat. No. 95-103 - EndoG Recombinant Protein

Species Reactivity :H, M, R

Short Description : Endonclease G

References
1.Li LY, Luo X, Wang X. Endonuclease G is an apoptotic DNase when released from mitochondria. Nature 2001; 412:95-9.
2.Parrish J, Li L, Klotz K, et al. Mitochondrial endonuclease G is important for apoptosis in C. elegans. Nature 2001; 412:90-4
3.Hengartner MO. Apoptosis. DNA destroyers. Nature 2001; 412:27, 29.
4.Widlak P, Li LY, Wang X, et al. Action of recombinant human apoptotic endonuclease G on naked DNA and chromatin substrates: cooperation with exonuclease and DNase I. J. Biol. Chem. 2001; 276:48404-9.