Trypsin is a porcine pancreas-derived enzyme that is commonly used for the dissociation and disaggregation of anchorage-dependent mammalian cells and tissues. The concentration of trypsin necessary to dislodge cells from their substrate is dependent primarily on the cell type and the age of the culture.
Trypsin-EDTA solutions are used to release adherent cells from culture surfaces. They are generally composed of variable concentrations of natural porcine pancreas-derived trypsin and EDTA. Trypsin 1X solutions can range from 0.025% to 0.5%. The reasons for the range of concentrations are differences in required trypsin activity or potency; differences in incubation times; and cell line susceptibility. The concentration of trypsin necessary to dislodge cells from their substrate is dependent primarily on the cell type and the age of the culture. Various formulations should be tested to determine the best product for a specific application.
Concentration: 2.5% (w/v)
Specific Activity: One USP trypsin unit is the activity causing a change in absorbance of 0.003 per minute under the conditions specified.
Suitability: Suitable for cell culture
Key Applications: Cell dissociation
Application Areas: Cell Counting, Viability, & Cryopreservation
Product Type: Reagents
Presentation: Clear, Colorless Solution
Format: Solution
Formulation: Potassium Chloride [KCl]: 400.0000 mg/L, Potassium Phosphate Monobasic [KH2PO4]: 60.0000 mg/L, Sodium Bicarbonate [NaHCO3]: 350.0000 mg/L, Sodium Chloride [NaCl]: 8000.0000 mg/L, Sodium Phosphate Dibasic [Na2HPO4]: 47.5000 mg/L, Dextrose: 1000.0000 mg/L, Phenol Red Sodium Salt: 17.0000 mg/L, Trypsin 1:250: 2500.0000 mg/L
pH: 5.0 - 6.0
Foreign activity: Mycoplasma, Porcine Parvovirus : Not detected
NOTES:
Osmolality: 310 - 480 mM/kg
Inhibitors: Trypsin is inhibited by organophosphorus compounds such as diisopropyl fluorophosphate and natural trypsin inhibitors from pancreas, soybean, lima bean and egg white. Silver ions are also potent inhibitors. Specific inhibitors are AEBSF, antipain, aprotinin, DFP, leupeptin, PMSF, TLCK, and Trypsin Inhibitor.
Composition: Trypsin is composed of two subunits, α-trypsin and β-trypsin. α-Trypsin is composed of two peptide chains and β-trypsin is composed of one chain.
Activity: One BAEE unit will produce a ΔA253 of 0.001 per minute at pH 7.6 at 25°C using BAEE as substrate. Reaction volume = 3.2 mL (1 cm light path). One TAME unit hydrolyzes 1 umole of p-toluene-sulfonyl-L-arginine methyl ester (TAME) per minute at 25°C, pH 8.2, in the presence of 0.001 M calcium ion.
Activity Conversion: 1 TAME unit = 19.2 USP or NF units = 57.5 BAEE
Units Specificity: The protease activity of trypsin is highly specific toward positively charged side chains with lysine and arginine. It forms complexes with α2-macroglobulin and can be used in the isolation of intact, detergent-free phycobilisomes and in the hydrolysis/condensation of carboxylic ester bonds.
Sterility: Sterile
Storage & Handling: Store between 0 and -20°C.