background:

Chymotrypsin, also known as chymotrypsin, belongs to the serine protease family and can specifically hydrolyze the peptide bonds formed by the carboxyl groups of aromatic amino acids. Chymotrypsinogen is secreted by the pancreas and enters the small intestine with pancreatic juice. It is cleaved by trypsin between Arg 15 and Ile 16 (numbered according to the Bovine chymotrypsin A sequence) into two parts connected by a disulfide bond. Subsequently, chymotrypsin cleaves the short peptide Ser 14-Arg 15 at Leu 13 and the short peptide Tyr 147-Asn 148 at Tyr 146 and Asn 148, forming three peptide chains connected by disulfide bonds and having biological activity.

Rhinogen ^® Chymotrypsin (Sequencing Grade) is a recombinant chymotrypsinogen obtained by recombinant expression in a recombinant Pichia yeast system and purification by multi-step chromatography. The active chymotrypsin obtained by activation (trypsin and autohydrolysis) has a purity of ≥95%, as shown in the figure below. Its amino acid sequence is completely consistent with the sequence of human chymotrypsin, and its molecular weight is about 25kDa. It can selectively hydrolyze the peptide bonds formed by the C-terminus of aromatic amino acids such as tyrosine, phenylalanine and tryptophan , and can also slowly hydrolyze the peptide bonds formed by leucine and methionine at a slower rate, and can also act on esters formed by sensitive amino acids.

Packing Specifications:

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

QIP-002 is provided as a lyophilized powder.

Product Source:

Rhinogen ^® Chymotrypsin (Sequencing Grade) is a recombinant chymotrypsin expressed in a recombinant Pichia pastoris system and purified by multi-step chromatography. Its amino acid sequence is completely consistent with that of human chymotrypsin, and its molecular weight is approximately 25kDa.

Product Quality:

The optimum pH is 7.0-9.5.

Storage conditions:

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

2. After dissolving with 50mM HAc , store at 2-8℃ and use within 2 days; or store the solution at -20℃ or below after aliquoting, with a validity period of 24 months. Avoid long-term storage at room temperature and repeated freezing and thawing.

Features:

Rhinogen ^® Chymotrypsin (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: the host protein residue is less than the limit requirement for biological products, and the purity is ≥95%;

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

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

4. High specific activity: specific activity (unit/mg) ≥2000 USP units/mg.

Application:

1. Proteomics research;

2. Peptide spectrum , peptide fingerprint or protein sequence analysis.

Recommended use:

1. Reagent preparation:

Dissolve Rhinogen ^{® Chymotrypsin (Sequencing Grade) dry powder} in 50mM HAc to a 1-10mg/ml solution.

2. Recommended usage:

In protein digestion, 50mM or 100mM NH ₄ HCO _{3 is used to dissolve the target protein, and then a certain amount of enzyme solution is added to make the enzyme amount: target protein (mass ratio) = 1: 50. The optimal reaction temperature is 30°C, and} the enzyme will self-cleave when the temperature exceeds 37°C .

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 human or animal diagnostic or therapeutic use.