|Place of Origin:||China, Shanghai|
|Certification:||NQA ISO 9001:2015|
|Minimum Order Quantity:||1U|
|Packaging Details:||1U/2U ice packing, carton|
|Delivery Time:||2-3 work days|
|Supply Ability:||1000,000g per month|
|Appearance:||White, Off-white Or Yellowish Powder||Specific Activity:||≥ 5.0 U/mg Pro.|
|Purity(SDS-PAGE):||Single Major Band||Molecular Weight(SDS-PAGE):||24.0±2.4 KDa|
|CAS No.:||66676-43-5||EC No.:||220.127.116.11|
glutamic acid protease,
High Stability Staphylococcus Aureus V8 Protease Without Miscellaneous Enzyme
Sequencing Grade V8 Protease
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. It can be used alone or in combination with other proteases when used in protein digestion, peptide profiling and peptide mass fingerprinting analysis. Recombinant V8 protease has activity between pH 4.0-10.0, and the optimum pH is 8.0-8.5. The inhibitors of V8 protease are diisopropylfluorophosphate (DFP), α2-macroglobulin and Nα-P-tosyl-L-lysine chloromethyl ketone (TLCK).
1. Product Information
Cat. NO.: V813
Full Name: Sequencing Grade Recombinant V8 Protease
Synonyms: V8 protease; Endoproteinase Asp-C; Endoproteinase Glu-C
Storage temperature: 2-8°C
Source: Recombinant V8 protease is a genetically engineered protein expressed in E.coli
2. MAIN FEATURES
|Source||Recombinant E. coli|
|Appearance||White or off white, or yellowish powder|
|Specific activity||≥ 5.0 U /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 in one minute at 25°C and pH 7.8 .
3. STABILITY OF STORAGE AND TRANSPORT
Storage stability: 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. There is no enzyme activity loss after 10 times repeated freezing and thawing.
Transportation stability: The product is stable by bue ice insulation transport.
4. RECOMMEND USAGE
Note that before using
1) Divide the material in according to amount used to avoid contamination or self-cutting.
2) Dissolve target protein.
The target protein is dissolved by 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), digesting the protein for 2-18h at 25-37 °C.
Table 1: Stability of V8 protease in the different concentrations of denaturant, urea 25 °C 2h, SDS 25 °C 12h
|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
Animal origin free: Recombinant V8 Protease has 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 quality of products.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 transport.
Compliance with regulatory requirements: Producing equipment and environment comply with relevant regulatory requirements, and the production process is in full compliance with NSF ISO 9001: 2015 quality system and GMP guidelines.
Complete quality documents: we can provide relevant regular support files in according to customers’ requirement.
Contact Person: Miss Eland