T5266

Trypsin from porcine pancreas

lyophilized powder, γ-irradiated, BioXtra, suitable for cell culture, 1,000-2,000 BAEE units/mg solid

• Numer CAS: 9002-07-7
• UNSPSC Code: 12352204
• NACRES: NA.75
• EC Number: 232-650-8
• MDL number: MFCD00082094
• Numer WE: 3.4.21.4 (BRENDA, IUBMB)
• Specific activity: 1,000-2,000 BAEE units/mg solid
• Biological source: Porcine pancreas
• Concentration: 25 mg/mL

Właściwości

• biological source: Porcine pancreas
• Quality Segment: 200
• sterility: γ-irradiated
• product line: BioXtra
• form: lyophilized powder
• specific activity: 1,000-2,000 BAEE units/mg solid
• mol wt: 23.8 kDa
• concentration: 25 mg/mL
• technique(s): cell culture | mammalian: suitable
• shipped in: wet ice
• storage temp.: 2-8°C

Opis

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 digestionsns†. 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.

Preparation Note

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

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. One BTEE unit = 320 ATEE units. Reaction volume = 3.2 mL (1 cm light path).

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.

Disclaimer

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.

Ta strona może zawierać tekst przetłumaczony maszynowo.

Informacja o środkach bezpieczeństwa

• Piktogramy: GHS08,GHS07
• Słowo ostrzegawcze: Danger
• Kody zagrożeń: H315,H319,H334,H335
• Ostrzeżenia: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Hazard Classifications: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• Narządy docelowe: Respiratory system
• Klasa składowania: 11 - Combustible Solids
• Co się dzieje?: WGK 1
• PPE (środki ochrony indywidualnej): dust mask type N95 (US), Eyeshields, Faceshields, Gloves