Staphylococcus Aureus v8 Protease,Sequencing Grade V8 Protease,a
genetically engineered protein expressed in E.coli.
Sequencing Grade V8 Protease
Full Name:Sequencing grade recombinant V8 protease;Staphylococcus aureus
Synonyms:V8 rotease;Endoproteinase Glu-C;Endoproteinase Asp-C
Storage temperature: 2-8°C
Source: Recombinant V8 protease is a genetically engineered protein
expressed in E.coli
Recombinant V8 protease is the serine protease family and can
specifically hydrolyzing glutamic acid or aspartic acid residues
carboxy side peptide bonds. The Glu carboxy terminal peptide bond
can be identified and cleaved in CH4COONH4 buffer (pH 7.8) and NH4HCO3 (pH 4.0 ).Identify and cleave the Glu or Asp carboxy terminal
peptide bond in phosphate buffer (pH 7.8),and the hydrolysis rate
of Glu was higher than that of Glu Asp. Recombinant V8 protease has
activity between pH 4.0-10.0, and the optimum pH is 8.0-8.5. It can
be used alone or in combination with other proteases when used in
protein digestion, peptide profiling and peptide mass
fingerprinting analysis. The inhibitors of V8 protease are
diisopropylfluorophosphate (DFP), α2-macroglobulin and
Nα-P-tosyl-L-lysine chloromethyl ketone (TLCK).
2. MAIN FEATURES
|Source||Recombinant E. coli|
|Appearance||White or off white, or yellowish powder|
|Specific activity||≥ 5.0 AU /mg pro|
|Purity(SDS-PAGE)||Single major band|
|Molecular Weight(SDS-PAGE)||24.0±2.4 kDa|
UNIT DEFINITION: One unit is defined as the amount of enzyme required to catalytic
substrate Z-Phe-Leu-Glu-4-nitranilide produced 1 μmol of
4-nitroaniline per minute at 25°C and pH 7.8 .
3. STABILITY OF STORAGE AND TRANSPORT
Stability of storage: Recombinant V8 protease should be stored
under 2-8°C in sealed container.It is stable within 6 months after
dissolved with1mM HCl and 50mM HAC at -20°C or below. It is stable
within 2 months after dissolved with 1mM HCl and 50mM HAC at
4°C.85% activity above can be kept after dissolved with 1mM HCl and
50mM HAC at 25°C or 37°C 12h. It is no activity loss after 10 times
repeated freezing and thawing.
Stability of transport: The product is stable by bue ice insulation transport.
4. RECOMMEND USAGE
Note that before using
1) Divide the material in according to usage quantity to avoid
pollution or self-cutting.
2) Dissolve target protein
The target protein is dissolved by cleavage buffer, for example
25-50 mM NH4HCO3 pH 7.8, if the solubility of target protein is not good, the
target protein will be denatured by adding urea, SDS, DTT or
heating. The effects of urea and SDS on V8 protease are shown in
the Table 1.
The recommended ratio of V8 protease and target protein is
1:20-1:100 (W/W).At 25 °C or 37 °C digest the protein 2-18h.
Table 1:Stability of V8 protease in the different concentrations of
denaturant, urea 25°C 2h, SDS 25°C12h
|denaturant||concentration||rest of activity|
|No any denaturant||0||100%|
4) Application example: recombinant V8 protease digests insulin
100 μl of insulin solution +198μl 0.4 M Tris-HCl pH 8.0 + 60μl 0.1%
V8 Protein solution + ultrapure water to 400μl. 37 °C water bath
reaction 2 h, plus 6μl phosphoric acid solution to terminate the
reaction, C18 column analysis:
A:Before digesting B:After digesting
Sequence grade recombinant protein: high specificity,good stability.
Animal origin free: Recombinant V8 Protease is no exogenous virus contamination,and
any animal origin material is not used in the production process.
Stable quality: Mass production can ensure stable and continuous batch
production.It is no difference between the batch and the product
quality is stable.
High purity: Higher specific activity.Host protein residues is less than the
limits of biological products.
Lyophilized powder: The product is lyophilized powder and is easy to store and
Compliance with regulatory requirements: Production equipment and
production environment comply with relevant regulatory
requirements, and the production process is in full compliance with
NSF ISO 9001: 2008 quality system and GMP guidelines.
Complete quality documents: we can provide relevant regular support files in according to
Sequencing grade trypsin
Sequencing grade chymotrypsin
Sequencing grade carboxypeptidase B