产品名称

Trypsin from porcine pancreas

一般描述

Trypsin consists of a single chain polypeptide of 223 amino acid residues, produced by the removal of the N-terminal hexapeptide from trypsinogen which is cleaved at the Lys - lle peptide bond. The sequence of amino acids is cross-ed by 6 disulfide bridges. This is the native form of trypsin, beta-trypsin. BETA-trypsin can be autolyzed, cleaving at the Lys - Ser residue, to produce alpha-trypsin. Trypsin is a member of the serine protease family.

含 量

27.0 - 33.0 mg/tab

CAS  NO.

9002-07-7

分子量

mol wt 23.8 kDa

MDL  number

MFCD00082094

适用范围

生物试剂

l  理化信息

外 观

白色到类白色圆形片剂

l  生物学信息

生物活性/药理作用

Trypsin cleaves peptides on the C-terminal side of lysine and arginine residues. The rate of hydrolysis of this reaction is slowed if an acidic residue is on either side of the cleavage site and  hydrolysis is stopped if a proline residue is on the carboxyl side of the cleavage site. The optimal pH for trypsin activity is 7-9. Trypsin can also act to cleave ester and amide ages of synthetic derivatives of amino acids. EDTA is added to trypsin solutions as a chelating agent that neutralizes calcium and magnesium ions that obscure the peptide bonds on which  trypsin acts. Removing these ions increases the enzymatic activity. Serine protease inhibitors, including DFP, TLCK, APMSF, AEBSEF, and aprotinin, amongst others, will inhibit Trypsin.

应 用

For use in immunohistochemical procedures to enhance staining and to unmask antigens after routine fixation and processing. For trypsin digestion of peptides, use a ratio of about 1:100 to 1:20 for trypsin:peptide. The typical use for this product is in removing adherent cells from a culture surface. The concentration of trypsin necessary to dislodge cells from their substrate is dependent primarily on the cell type and the age of the culture. Trypsins have also been used for the re-suspension of cells during cell culture, in proteomics research for digestion of proteins and in various in-gel digestionsns†. Additional applications include assessing crystallization by membrane-d techniques and in a study to determine that protein folding rates and yields can be limited by the presence of kinetic traps.

样品准备说明

This product is a lyophilized powder soluble in Hank′s Balanced Salt Solution at 25mg/mL. For applications that require EDTA, solubilizing trypsin should be done with a buffered salt solution contaiing no Ca₂₊ or Mg₂₊.

注 意

Solutions in 1mM HCl are stable for 1 year in aliquots and stored at -20°C. The presence of Ca₂₊ will also diminish the self-autolysis of trypsin and maintain its stability in solution. Trypsin will also retain most of its activity in 2.0M urea, 2.0M guanidine HCl, or 0.1% (w/v) SDS.

有效单位定义

One BAEE unit will produce a ΔA²⁵³  of 0.001 per min at pH7.6 at 25°C using BAEE as substrate. One BTEE unit = 320 ATEE units. Reaction volume = 3.2mL (1cm light path).

l  存储

存储温度

-20°C

l  注意事项及免责声明

本产品仅用于实验研究，不得作为药物使用，不得用于家用或其它用途。

l  参考文献

1.    http://www.drugbank.ca

2.    https://ncit.nci.nih.gov

3.    https://www.ncbi.nlm.nih.gov