| Assay Method Information | |
| | Assay for the Determination of the Capacity to Inhibit Calpain 1 |
| Description: | Activity of Calpain 1 was measured by using Calpain-Glo Protease kit from Promega. Assays were run in 384-well black plates with clear bottom. 5 μl of compounds in a 10 mM HEPES and 1 mM EDTA aqueous solution were incubated with 15 μL of a solution of 16 ng/mL calpain 1 (Sigma Aldrich) for 2 minutes and then 20 μL of Calpain-Glo Reagent provided in the kit were added to each well. Luminiscence elicited by calpain 1-induced substrate lysis was measured in a Hamamatsu FDSS 7000 reader and data was calculated from the maximal luminescence peak. Calpain inhibition was calculated from the formula:% inhibition=(LT−LB)*100/(LC−LB);wherein LT is the luminescence observed in the tested compound, LB is blank luminescence obtained from a well without calpain 1 and LC is the luminescence observed in control wells including calpain1 in the absence of inhibitor. |
| Affinity data for this assay | |
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