Endoproteinase Glu-C,specification,price,image-Bio-Equip in China

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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. [2] Prasad L , Leduc Y , Hayakawa K ,et al. The structure of a universally employed enzyme: V8 protease from Staphylococcus aureus[J].Acta Crystallographica Section D Biological Crystallography, 2004.

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.