l General Information |
Product Name | Trypsin from bovine pancreas |
Assay | ≥9,000 BAEE units/mg protein; protein, ≥80% | CAS Number | 9002-07-7 |
Molecular Weight | mol wt 23.8 kDa | MDL number | MFCD00082094 |
Suitability | BioReagent, suitable for cell culture |
l Physical and Chemical Information |
Appearance | essentially salt-free,White lyophilized powder |
Solubility(25°C) | hydrochloric acid: soluble 1 mM |
l Biological Information |
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 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. |
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. |
Preparation Note | This product is from pancreas sourced from New Zealand. It is soluble in 1 mM HCl at 1 mg/mL. For applications that involve EDTA, solubilizing trypsin should be done with a buffered salt solution containing no Ca2+ or Mg2+. |
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. |
Unit Definition | One BAEE unit will produce a A253of 0.001 per minute at pH 7.6 at 25°C using BAEE as a substrate. One BTEE unit = 320 ATEE units |
l Storage |
Storage temp. | -20°C |
l Precautions and Disclaimer |
This product is for R&D use only, not for drug, household, or other uses. |
l References |
1. http://www.drugbank.ca 2. https://ncit.nci.nih.gov 3. https://www.ncbi.nlm.nih.gov |
