Recombinant protein corresponding to Ala16-Ser247 from human Trypsin 2 expressed in NSO cells.
Applications: Western Blot: 0.1-0.2ug/ml with the appropriate secondary reagents to detect human Trypsin 1/2/3. The detection limit for rhTrypsin 1/2/3 is approximately 1ng/lane under non-reducing and reducing conditions. Immunoprecipitation: Used to immunoprecipitate rhTrypsin 1/2/3 from conditioned media of transfected NS0 cells. Neutralization: Used to inhibit/neutralize the enzymatic activity of rhTrypsin 2. The antibody was preincubated with the enzyme at different molar ratios at room temperature for 30 minutes. The enzyme was then assayed with a fluorogenic peptide substrate. 50% of the proteolytic activity was inhibited by the antibody at approximately 0.3ug/ml (IC50) under conditions in which the enzyme was present at 0.02ug/ml and the substrate concentration was 10uM. Considering the molecular masses of the enzyme (26 kD) and the antibody (150 kD), IC50 was achieved at approximately 3:1 molar ratio of the antibody to the enzyme. The antibody did not inhibit the activity of rhTrypsin 1 and rhTrypsin 3 significantly. Direct ELISA: 0.5-1.0ug/ml with the appropriate secondary reagents to detect human Trypsin 1/2/3. The detection limit for rhTrypsin 1/2/3 is approximately 0.8ng/well. Optimal dilutions should be determined by each laboratory for each application. Storage and Stability: Lyophilized powder may be stored at 4C for short-term only. Reconstitute to nominal volume by adding sterile PBS and store at -20C. Reconstituted product is stable for 12 months at -20C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
