background:

Endoproteinase Glu-C (also known as V8 protease, glutaminase, asparaginase, Staphylococcus aureus V8 protease) belongs to the serine protease family and can specifically hydrolyze the carboxyl terminal peptide bond of glutamic acid (Glu) or aspartic acid (Asp) residues. Its specificity is directly affected by the buffer components. It recognizes and cuts the carboxyl terminal peptide bond of Glu in NH4HCO3 buffer at pH 7.8 and CH3COONH4 buffer at pH 4.0, and recognizes and cuts the carboxyl terminal peptide bond of Glu or Asp in phosphate buffer at pH 7.8, and the hydrolysis rate of Glu is higher than that of Asp.

Rhinogen ^® Glu-C (Sequencing Grade) is a recombinant protease expressed by a recombinant E. coli system and purified by multi-step chromatography. It has a molecular weight of about 24kD and can be used alone or in combination with other proteases for protein restriction sequencing, peptide mapping and peptide fingerprinting . It is active between pH 4.0-9.0, with an optimum pH of 8.0-8.5. Inhibitors of V8 protease include diisopropyl fluorophosphate (DFP), α2-macroglobulin and Nα-P- tosyl -L-lysine chloromethyl ketone (TLCK).

SHAPE \* MERGEFORMAT

Packing Specifications:

Rhinogen ^® Trypsin (Sequencing Grade) packaging specifications are as follows:

QIP-005 is provided as a lyophilized powder.

Product Source:

Rhinogen ^® Glu-C (Sequencing Grade) is a recombinant protease expressed in a recombinant E. coli system and purified by multi-step chromatography. Its molecular weight is approximately 24kDa.

Product Quality:

SDS-PAGE analysis showed purity ≥95%; no contaminating protease activity was detected.

Product Features:

It is active between pH 4.0-9.0, with the optimum pH being 8.0-8.5.

Storage conditions:

1. Use ice packs for transportation. Please place the enzyme at -20℃ immediately after receiving the product and seal it to prevent moisture.

2. After dissolving in water or 50mM pH8.0 Tris-HCl to 0.5mg/ml, store below -20℃ and it is stable for 6 months; store at 2-8℃ (sterile), it is stable for 2 months; the activity can be maintained above 85% at 25℃ or 37℃ for 12h; avoid leaving it at room temperature for a long time, it is recommended to store it in aliquots to avoid repeated freezing and thawing.

Features:

Rhinogen ^® Glu-C (Sequencing Grade) is a highly purified and very stable recombinant protease with high stability and high specific activity. It is suitable for peptide mapping , peptide fingerprinting or protein sequence analysis in proteomics research.

1. High purity: no contaminating protease, purity ≥ 95%;

2. High stability: Each batch of products undergoes strict quality control to achieve batch-to-batch stability ;

3. No animal origin: Recombinant production, no exogenous virus contamination, and no animal origin raw materials are used in the production process .

application:

1. Peptide spectrum , peptide fingerprint or protein sequence analysis;

2. Proteomic research.

Recommended use:

1. Reagent preparation:

1) Use water or 50mM pH8.0 Tris-HCl to dissolve the dry powder to obtain a Glu-C solution with a concentration of 0.5mg/ml-1mg/ml. After dissolution, it is recommended to divide it according to the single use amount to minimize contamination or self-cutting.

2. Recommended usage:

1) Dissolve the target protein: Use enzyme digestion buffer (such as 25-50mM pH7.8 NH ₄ HCO ₃ buffer) to dissolve the target protein. If the solubility of the target protein is low, you can consider denaturing the target protein (such as adding urea, SDS, DTT or heating). The effects of urea and SDS on Glu-C protease are shown in the table below:

2) Target protein cleavage: The recommended usage of Glu-C protease is Glu-C protease: target protein = 1:20-1:100 (w/w), the optimal pH is 8.0-8.5, and the enzyme cleavage is at 25℃ or 37℃ for 2-18hrs.

Note: For different protein samples, it is necessary to experiment to find the most appropriate enzyme concentration and reaction time.

Instructions:

This product is for research use only and is not intended for diagnostic or therapeutic use in humans or animals.