| Assay Method Information | |
| | α-Chymotrypsin Inhibition Assay |
| Description: | The α-chymotrypsin inhibition activity was evaluated in 50 mM Tris-HCl buffer pH 7.6 with 10 mM CaCl2. α-Chymotrypsin (bovine pancreas) at the final concentration of 12 units/mL (prepared in buffer mentioned above) with the various concentrations of test compounds, prepared in DMSO, was incubated at 30 °C for25 min. The reaction was started by the addition of the substrate, N-succinyl-L-phenylalanine-p-nitroanilide (SPpNA; 0.4 mM final concentration prepared in the buffer as above). The change in absorbance by released p-nitroaniline was continuously monitored at 410 nm [Cloudhary et al., Phytochem. Lett., 4:404-406]. All the experiments were performed in triplicate in a final volume of 200 μL using a micro-plate reader (SpectraMax M2, Molecular Devices, CA, USA). |
| Affinity data for this assay | |
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