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V8 Protease
V8 Protease
Place of Origin:
China
Brand:
YaxinBio
Model:
V813
Price:
65$/U
Hits:
2859 
Updated:
10/10/2024
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    Sequencing Grade V8 Protease

    Catalog Number:V813

    CAS:66676-43-5

    EC:3.4.21.19

    Full Name:,Sequencing grade recombinant V8 protease;Staphylococcus aureus strain V8

    Synonyms:V8 protease;Endoproteinase Glu-C;Endoproteinase Asp-C 

    Storage temperature: 2-8°C

    Source: Recombinant V8 protease is a genetically engineered protein expressed in E.coli 

     

    1. DESCRIPTION    

    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.

    SPECIFIC ACTIVITY: At 25°C, the activity measured with Z-Phe-Leu-Glu-4-nitranilide as a substrate is about 20U/mg pro. (Corresponding to the activity measured with casein is about 500 U / mg pro.at 37°C).

     

    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 blue 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.

    3) digest

       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%

    urea

    2M

    95%

    5M

    12%

    8M

    0

    SDS

    0.1%

    90%

    0.5%

    80%

    1%

    50%

     

    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

    5.PRODUCT ADVANTAGE

    • 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 transport.
    • 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 customers’ requirement.

    6.RELATED PRODUCTS

    Recombinant enterokinase

    Sequencing grade trypsin

    Sequencing grade chymotrypsin

    Sequencing grade carboxypeptidase B

    bio-equip.cn
    Shanghai Yaxin Biotechnology Co. Ltd. is a biotech company founded in 2008 that focuses on the research and the production of recombinant proteins. Shanghai YaxinBio,located in Shanghai Juke Biotech Park, is ISO 9001 registered company since 2009.


    Our main products are Animal Origin Free recombinant carboxypeptidase B and recombinant trypsin, which are of high importance for the production of human recombinant insulin.YaxinBio recombinant carboxypeptidase B and recombinant trypsin have no trace of other contaminating enzyme activities or enzyme inhibitors. They have been successfully used in the production process of recombinant insulin by several companies. Recombinant Trypsin can also be used in the process of cell culture.


    Shanghai YaxinBio continuously strives to bring to its worldwide customers the best recombinant protein products. We will continue strengthen our product management and our internal system to meet the demands of biotech companies and scientists in the field of life sciences.

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