Assay Method Information |
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| m-Calpain Inhibition Assay |
Description: | Inhibition of m-calpain was assayed in the reaction buffer (50 mM Tris-HCl, 100 mM NaCl, 5 mM β-mercaptoethanol, and 1 mM EDTA, pH 7.5) with 50 µM substrate B, 5 mM CaCl2 and 2.88 U/mL m-calpain. The reaction was initiated by sequential addition of substrate B, m-calpain, each compound, and finally a CaCl2 solution. The reaction mixture (100 µL) was incubated at room temperature for 30 min with shaking. Fluorescence intensities were measured at 320 nm excitation and 420 nm emission wavelengths. MDL28170 was used as a positive control for µ- and m-calpain inhibition. |
Affinity data for this assay | |
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