Trypsin from bovine pancreas
SIGMA/T8802 - TPCK Treated, essentially salt-free, lyophilized powder, ≥10,000 BAEE units/mg protein
CAS Number: 9002-07-7
EC Number: 232-650-8
MDL Number: MFCD00082094
Product Type: Chemical
Enterokinase activates pancreatic trypsinogen to trypsin by the hydrolysis of a hexapeptide(for bovine trypsin at the Lys6 - Ile7 peptide bond) from the NH2 terminus. Bovine trypsinogen consists of a single polypeptide chain of 229 amino acids and is cross linked by six disulfide bridges. Trypsin can autocatalytically activate more trypsinogen to trypsin. Trypsin consists of a single chain polypeptide of 223 amino acid residues. This native form of trypsin is refered to as β-trypsin. Autolysis of β-trypsin (which is cleaved at Lys131- Ser132 in the bovine sequence) results in α-trypsin which is held together by disulfide bridges.
| biological source | bovine pancreas |
| composition | protein, ≥95% |
| foreign activity | Chymotrypsin ≤0.1 BTEE units/mg protein |
| form | essentially salt-free, lyophilized powder |
| mol wt | 23.8 kDa |
| Quality Level | 200  |
| solubility | hydrochloric acid: soluble 1 mM, clear |
| specific activity | ≥10,000 BAEE units/mg protein |
| sterility | aseptically filled |
| storage temp. | −20°C |
| Analysis Note: | Protein determined by E |
| 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. |
| Application: | Trypsin can be used to release adherent cells from tissue culture plates for passaging. Trypsin has been used in a study to assess the effects of macromolecular crowding on the structural stability of human α-lactalbumin. Trypsin has also been used in a study to investigate BN-PAGE analysis of Trichoderma harzianum secretome. |
| 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. |
| General description: | The trypsin molecule has two domains: one is related to the enzyme active site and the tryptophan residues; the other is related to the 8-anilinonaphthalene-1-su |
| Other Notes: | One BAEE unit will produce a ΔA253 of 0.001 per min at pH 7.6 at 25 °C using BAEE as substrate. Reaction volume = 3.2 ml (1 cm light path). |
| Other Notes: | One BAEE unit will produce a A253 of 0.001 per minute at pH 7.6 at 25°C using BAEE as a substrate. |
| Other Notes: | View more information on trypsin at www.sigma-aldrich.com/enz |
| Preparation Note: | TPCK treated |
| Symbol |   GHS07,GHS08 |
| Signal word | Danger |
| Hazard statements | H315 - H319 - H334 - H335 |
| Precautionary statements | P261 - P264 - P271 - P302 + P352 - P304 + P340 + P312 - P305 + P351 + P338 |
| Hazard Codes | Xn |
| Risk Statements | 36/37/38-42 |
| Safety Statements | 22-24-26-36/37 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 1 |
| activity | specific activity: ≥10,000 BAEE units/mg protein |
| Storage Temp. | −20°C |
| Enzyme Commission (EC) Number | 3.4.21.4 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352204 |