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 Norgen’s proprietary resin as the separation matrix. Briefly, trypsin is added to the protein sample and the sample is then loaded onto Norgen'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 ProteoSpin^TM 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 convenient 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.