The Thermo Scientific Pierce V-8 Protease Kit contains Staphylococcus aureus V-8 protease immobilized on resin and three digestion buffers for specific cleavage at glutamic acid residues or cleavage at glutamic and aspartic acid residues.
There are two major problems with any proteolytic digestion. The first problem is that the reaction must be quenched to stop the digestion, thus diluting out the sample. The second problem is that the sample is contaminated with the protease and possibly autodigested protease fragments. These problems are surmounted with immobilized V-8 protease. With immobilized proteases, no quenching is necessary and there are no problems with protease contamination of the sample or autodigestion of the protease. Simply separate the sample solution from the agarose matrix and these problems are eliminated.
- Specificity for glutamic acid is achieved in ammonium bicarbonate, pH 7.8 or ammonium acetate, pH 4
- Enzyme is active in the presence of many denaturing agents such as SDS, urea and guanidine-HCl
- Immobilized enzyme eliminates protease contamination in samples
- Three V-8 digestion buffers in the kit allow specificity for glutamate or glutamate and aspartate residues
Staphylococcus aureus V-8 protease, also known as endoproteinase Glu-C, has two pH optima, one at pH 4.0 and the other at pH 7.8, and it is specific for cleavage at the carboxy terminus of glutamic acid and aspartic acid residues. The protease is further specific for only glutamic digestion in buffers which do not contain phosphate at either pH optimum. Phosphate buffers at pH 7.8 will facilitate digestion at both glutamic and aspartic acid residues.
S. aureus V-8 protease protease is very stable, being able to retain its activity even in 6 M urea, 5.5 M guanidine•HCl, 0.5% SDS, or 0.5% SDS + 6 M urea at 0°C, pH 7.6. This is related to the fact that the native structure of the protease is stabilized mainly by electrostatic interactions, and not by hydrogen bonding and ß-structures.
Properties of V-8 Protease.
||endoproteinase Glu-C, Glu-C protease, staphylococcal serine proteinase, Staphylococcus V8 serine endopeptidase
||Cleaves specifically at the carboxyl side of aspartate and glutamate residues
||Staphylococcus aureus , V-8 strain
||Specific protein fragmentation for protein identification and structure analysis
||pH 4 or pH 7.8, 37°C
||Store at 4°C
||PMSF, DFP, Aprotinin
- Drapeau, G.R., et al. (1972). J. Biol. Chem. 247, 6720.
- Houmard, J. and Drapeau, G. (1972). Proc. Natl. Acad. Sci. USA 69(12), 3506-3509.
- Geiben-Lynn, R., et al. (2002) J. Biol. Chem.277, 42352-42357.
Pierce Trypsin, MS Grade (Part No. 90057)
MS-Grade Proteases and Digestion Kits for Mass Spectrometry
Standard-Grade Proteases and Protease Resins
Enzymes for Cell Lysis