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T0303

Sigma-Aldrich

Trypsin from porcine pancreas

Type IX-S, lyophilized powder, 13,000-20,000 BAEE units/mg protein

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Synonym(s):
Serine Protease 1
CAS Number:
Enzyme Commission number:
EC Number:
MDL number:

type

Type IX-S

Quality Level

form

lyophilized powder

specific activity

13,000-20,000 BAEE units/mg protein

mol wt

23.8 kDa

application(s)

diagnostic assay manufacturing

foreign activity

chymotrypsin ≤1.0 units/mg protein

storage temp.

−20°C

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This Item
T1426T4799T5266
Trypsin from porcine pancreas Type IX-S, lyophilized powder, 13,000-20,000 BAEE units/mg protein

T0303

Trypsin from porcine pancreas

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

T1426

Trypsin from bovine pancreas

Trypsin from porcine pancreas lyophilized powder, BioReagent, suitable for cell culture, 1,000-2,000 BAEE units/mg solid

T4799

Trypsin from porcine pancreas

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

T5266

Trypsin from porcine pancreas

form

lyophilized powder

form

essentially salt-free, lyophilized powder

form

lyophilized powder

form

lyophilized powder

application(s)

diagnostic assay manufacturing

application(s)

diagnostic assay manufacturing

application(s)

-

application(s)

-

storage temp.

−20°C

storage temp.

−20°C

storage temp.

−20°C

storage temp.

2-8°C

mol wt

23.8 kDa

mol wt

23.8 kDa

mol wt

23.8 kDa

mol wt

23.8 kDa

foreign activity

chymotrypsin ≤1.0 units/mg protein

foreign activity

Chymotrypsin ≤0.1 BTEE units/mg protein

foreign activity

-

foreign activity

-

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.

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.

Unit Definition

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).

Preparation Note

This product is a lyophilized powder soluble in Hank′s Balanced Salt Solution at 25 mg/mL.

pictograms

Exclamation markHealth hazard

signalword

Danger

Hazard Classifications

Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3

target_organs

Respiratory system

Storage Class

11 - Combustible Solids

wgk_germany

WGK 1

ppe

dust mask type N95 (US), Eyeshields, Faceshields, Gloves


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Protocols

Continuous spectrophotometric rate determination method using BAEE substrate measures trypsin activity, essential for enzyme characterization.

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