The sequence coverage of Bovine Serum Albumin samples was 98.3% after Glu-C enzymolysis and LC-MS/MS analysis
Endoproteinase Glu-C
Endoproteinase Glu-C
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$156 USD
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Product Details
Product Details
Product Specification
| Species | Staphylococcus aureus |
| Synonyms | Glutamyl endopeptidase、V8 protease、Endoproteinase Glu-C、V8 proteinase、V8、Glu-C |
| Expression System | E.coli |
| Molecular Weight | 25kDa (Reducing) |
| Purity | ＞95% by SDS-PAGE |
| Tag | His Tag |
| Physical Appearance | Lyophilized Powder |
| Reconstitution | To get the most use out the enzyme, resuspend the enzyme in 500 μl H2O and aliquot 50 ul each in 10 tubes. |
| Stability & Storage | Store at -25 ~ -15℃ for 2 years |
| Reference | [1] Yabuta M , Ochi N , Ohsuye K .Hyperproduction of a recombinant fusion protein of Staphylococcus aureus V8 protease in Escherichia coli and its processing by OmpT protease to release an active V8 protease derivative.[J].Appl Microbiol Biotechnol, 1995, 44(1-2):118-125. |
Background
Staphylococcus aureus V8 protease (Endoproteinase Glu-C) belongs to the serine protease family and can specifically hydrolyze carboxy-terminal peptide bonds of glutamic acid (Glu) or aspartate (Asp) residues. Its specificity is directly affected by the composition of the buffer, recognizing and cutting Glu carboxy-terminal peptide bonds in NH4HCO3 of pH7.8 and CH3COONH4 of pH4.0, recognizing and cutting Glu or Asp carboxy-terminal peptide bonds in phosphate buffers of pH7.8，but the hydrolysis rate of Glu was higher than that of Asp. Glu-C has been reported to be active in 0.2% SDS (sodium lauryl sulfate) and 4.0M urea.
Components
Supplied in dry format from Tris-HCl and sodium chloride buffer
Protocol
For the digestion of peptides or proteins, the ratio of enzyme to substrate between is 1:20 and 1:100 (w/w) for recommended. Dissolve the peptide or protein and was enzymolysis in 100 mM NH4HCO3, pH 7.8 or 100 mM Tris-HCl, pH 7.8, the recommended incubation time is 2-18 hours at 37°C, depending on the temperature and the ratio of enzyme to substrate.
Guidelines
Repeated freeze/thawing is not recommended.
Unit Definition
Using 2-Ph-leu-Glu-4-Na as substrate, the amount of enzyme required to produce 1μmol 4-Nitroaniline in 1 minute at pH7.8, 25℃ was 1unit.
Picture
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Bioactivity
SDS-PAGE
2μg (R: reducing condition, N: non-reducing condition).
