T8802

Trypsin from bovine pancreas

TPCK Treated, essentially salt-free, lyophilized powder, ≥10,000 BAEE units/mg protein

• CAS Number: 9002-07-7
• UNSPSC Code: 12352204
• NACRES: NA.54
• EC Number: 232-650-8
• MDL number: MFCD00082094
• EC Number: 3.4.21.4 (BRENDA, IUBMB)
• Specific activity: ≥10,000 BAEE units/mg protein
• Biological source: bovine pancreas

Properties

• biological source: bovine pancreas
• Quality Level: 200
• sterility: aseptically filled
• form: essentially salt-free, lyophilized powder
• specific activity: ≥10,000 BAEE units/mg protein
• mol wt: 23.8 kDa
• composition: protein, ≥95%
• solubility: hydrochloric acid: soluble 1 mM, clear
• foreign activity: Chymotrypsin ≤0.1 BTEE units/mg protein
• storage temp.: −20°C

Description

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-sulfonate binding.

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.

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. [1] Trypsin has also been used in a study to investigate BN-PAGE analysis of Trichoderma harzianum secretome. [2]

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

TPCK treated

Analysis Note

Protein determined by E1%/280

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).

One BAEE unit will produce a A₂₅₃ of 0.001 per minute at pH 7.6 at 25°C using BAEE as a substrate.

View more information on trypsin at www.sigma-aldrich.com/enzymeexplorer

Safety Information

• pictograms: GHS08,GHS07
• signalword: Danger
• hcodes: H315,H319,H334,H335
• pcodes: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Hazard Classifications: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• target_organs: Respiratory system
• Storage Class: 11 - Combustible Solids
• wgk: WGK 1
• ppe: dust mask type N95 (US), Eyeshields, Faceshields, Gloves