On-Column Proteolytic Digestion Kit

On-Column Proteolytic Digestion Kit

Catalog# : NB-17500
Price/Unit : $220.00
Unit : 25 preps
For the rapid and convenient on-column digestion of proteins

* Fast and simple procedure to generate digested peptides
* Simultaneous digestion and concentration
* Peptides are ready for applications such as mass spectrometry and SDS-PAGE

The ProteoSpin On-Column Proteolytic Digestion kit is highly efficient in the enzymatic digestion of simple and complex protein samples using trypsin, and the subsequent purification of the resulting peptides.

Purification is based on spin column chromatography using Bio-Synthesis's proprietary resin as the separation matrix. Briefly, trypsin is added to the protein sample and the sample is then loaded onto Bio-Synthesis's column (BIND). Binding occurs if the protein or proteins of interest retain a net positive charge at the binding pH. At the binding pH, the activity of trypsin is inhibited. Non-specifically bound materials such as salts are then washed from the column and the bound protein and enzyme remain (WASH). The trypsin is then activated and the digestion proceeds for approximately 2 hours. The peptides are then eluted into a small volume (ELUTE). Please see the procedure flowchart to the right.

Each column in the ProteoSpinTM On-Column Proteolytic Digestion Kit is able to process, digest and purify samples containing up to 15 µg of protein. The peptides generated are complete, with no additional artifacts being detected in mass spectrometry. The simultaneous protein digestion and volumetric concentration of the purified peptides makes the kit a cenvenient method for preparing peptides to be analyzed by many downstream applications.

FEATURES AND BENEFITS

* Fast method for generating peptides - Compared to traditional methods of peptide generation and purification, the ProteoSpin™ On-Column Proteolytic Digestion kit is very rapid.
* Does not generate artifacts - Peptide generation is complete, with no generation of additional artifacts being detected in mass spectrometry.
* Digestion and purification in a single column - The use of multiple columns and tubes for digestion and purification is avoided; lower cost and increased convenience for user.

APPLICATIONS
The simultaneous protein digestion and volumetric concentration of the purified peptides makes the kit a convenient method for preparing peptides to be analyzed by many downstream applications such as:

* SDS-PAGE
* 2D gel electrophoresis
* MALDI-TOF
* LC/MS
* LC/MS/MS

Figure 1. Generation of Tryptic BSA Peptides without the Generation of Artifacts - MALDI-TOF spectra of the peptides that resulted after BSA was digested on-column with trypsin using the ProteoSpin™ On-Column Proteolytic Digestion Kit (Panel A) or digested in a typical liquid environment (Panel B). For both digestions, 15 g of BSA was used as the input and 0.3 µg of Trypsin was used for the digestion. For on-column digestion, the 15 µg of BSA was loaded onto the column with the trypsin, the BSA was washed, then the on-column digestion proceeded and the resulting peptides were eluted. By comparing the spectra, it can be seen that the ProteoSpin™ kit (Panel A) resulted in the generation of the same peptide profile in MALDI-TOF as the liquid digestion did (Panel B). Furthermore, no artifacts were found to arise from the use of the column, and all the same peaks could be detected in both spectra.

On-Column Proteolytic Digestion Kit Contents
1. Column Equilibration and Wash Buffer
2. Column pH Binding Buffer
3. Column Activation Buffer
4. Micro Spin Columns (25)
5. Elution Tubes (25)
6. Product Insert
Long-Term Storage : All solutions should be kept tightly sealed and stored at room temperature. Once opened, the solution should be stored at 4°C.
Product Usage
For laboratory research use only, not for diagnostic use! Not for use in humans.