T4799

Trypsin from porcine pancreas

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

• Szinonimák: Cocoonase, Tryptar, Tryptase
• CAS-szám: 9002-07-7
• Enzyme Commission szám: 3.4.21.4 (BRENDA, IUBMB)
• EC-szám: 232-650-8
• MDL-szám: MFCD00082094
• UNSPSC kód: 12352204
• NACRES: NA.75

Tulajdonságok

• biológiai forrás: Porcine pancreas
• Minőségi szint: 200
• termékcsalád: BioReagent
• Forma: lyophilized powder
• specifikus aktivitás: 1,000-2,000 BAEE units/mg solid
• molekulatömeg: 23.8 kDa
• koncentráció: 25 mg/mL
• technika/technikák: cell culture | mammalian: suitable; single cell analysis: suitable
• pH: 7.6
• kiszállítva: ambient
• tárolási hőmérséklet: −20°C

Leírás

Általános leírás

Trypsin is applicable for tissue disaggregation, due to its effective action and tolerance towards different cell type and serum-induced neutralization.

Alkalmazás

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.

Trypsin from porcine pancreas has been used to digest chicken bones. It has also been used in the isolation of luteal endothelial cells.

Biokémiai/fiziológiai hatások

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.

Komponensek

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.

Vigyázat

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.

Egység definíció

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).

Elkészítési megjegyzés

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

Biztonsági adatok

• Piktogramok: GHS08,GHS07
• Figyelmeztetés: Danger
• Figyelmeztető mondatok: H315,H319,H334,H335
• Óvintézkedésre vonatkozó mondatok: P261 - P264 - P271 - P280 - P302 + P352 - P305 + P351 + P338
• Veszélyességi osztályok: Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
• Célzott szervek: Respiratory system
• Tárolási osztály kódja: 11 - Combustible Solids
• WGK: WGK 1
• Egyéni védőeszköz: dust mask type N95 (US), Eyeshields, Faceshields, Gloves