性能描述

胰蛋白酶经TPCK处理，因此，胰蛋白酶活力更高，
糜胰蛋白酶活性被抑制。TPCK-胰蛋白酶能够专一性地切割蛋白
分子中精氨酸和赖氨酸位点，是蛋白质肽段分析重要工具酶之
一。
用于梅切或蛋白测序分析的工作条件
缓冲液：50mM NH4HCO3 (pH 7.8)
用量：TPCK-TRYPSIN：目标蛋白＝1:20-50
反应条件：37℃，2-3 小时

form    essentially salt-free, lyophilized powder 
mol wt    mol wt 23.8 kDa 
Market Segment    Diagnostic Assay Manufacturing 
foreign activity    Chymotrypsin ≤0.1 BTEE units/mg protein 
storage temp.    −20°C 
Components
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-linked 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.

Caution
Solutions in 1 mM 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 2.0 M urea, 2.0 M guanidine HCl, or 0.1% (w/v) SDS.

Preparation Note
Solubilizing trypsin should be done with a buffered salt solution containing no Ca2+ or Mg2+. This product is from pancreas sourced from New Zealand. It is soluble in 1 mM HCl at 1 mg/mL.
It is also TPCK-treated and dialyzed. Treatment with L-1-Tosylamide-2-phenyleth​yl chloromethyl ketone (TPCK) reduces the chymotrypsin activity which is usually present in trypsin.

Application
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 digestions. Additional applications include assessing crystallization by membrane-based techniques and in a study to determine that protein folding rates and yields can be limited by the presence of kinetic traps.

Biochem/physiol Actions
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 linkages 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.