T5266
Trypsin from porcine pancreas
lyophilized powder, γ-irradiated, BioXtra, suitable for cell culture, 1,000-2,000 BAEE units/mg solid
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About This Item
Polecane produkty
pochodzenie biologiczne
Porcine pancreas
Poziom jakości
sterylność
γ-irradiated
linia produktu
BioXtra
Postać
lyophilized powder
aktywność właściwa
1,000-2,000 BAEE units/mg solid
masa cząsteczkowa
23.8 kDa
stężenie
25 mg/mL
metody
cell culture | mammalian: suitable
Warunki transportu
wet ice
temp. przechowywania
2-8°C
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Zastosowanie
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.
Działania biochem./fizjol.
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.
Serine protease inhibitors, including DFP, TLCK, APMSF, AEBSEF, and aprotinin, amongst others, will inhibit Trypsin.
Komponenty
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.
Przestroga
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.
Definicja jednostki
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).
Uwaga dotycząca przygotowania
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+.
For applications that involve EDTA, solubilizing trypsin should be done with a buffered salt solution contaiing no Ca2+ or Mg2+.
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Hasło ostrzegawcze
Danger
Zwroty wskazujące rodzaj zagrożenia
Zwroty wskazujące środki ostrożności
Klasyfikacja zagrożeń
Eye Irrit. 2 - Resp. Sens. 1 - Skin Irrit. 2 - STOT SE 3
Organy docelowe
Respiratory system
Kod klasy składowania
11 - Combustible Solids
Klasa zagrożenia wodnego (WGK)
WGK 1
Środki ochrony indywidualnej
dust mask type N95 (US), Eyeshields, Faceshields, Gloves
Certyfikaty analizy (CoA)
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Protokoły
This procedure is for products with a specification for Trypsin activity using Na-Benzoyl-L-arginine ethyl ester (BAEE) as a substrate. The procedure is a continuous spectrophotometric rate determination (A253, Light path = 1 cm).
Ciągła spektrofotometryczna metoda oznaczania szybkości przy użyciu substratu BAEE mierzy aktywność trypsyny, niezbędną do charakterystyki enzymu.
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