| Assay Method Information | |
| | Assay of Inhibitory Effect on Cardiac yosin ATPase acAtivity |
| Description: | a) 250 μL of ice-cold PM12 buffer was added to 250 μg of S1 myosin to a protein concentration of 1 mg/mL.b) The reagents were successively added in the following order and mixed to give a reaction mixture:400 μL of PM12,400 μL of 5× MSEG (from the ATPase assay biochem kit),1200 μL of actin/cardiac tropomyosin/troponin complex,40 μL of myosin S1,40 μL of 100× PNP (from the ATPase assay biochem kit),10.4 μL of 100 mM ATP.c) 10 μL of 440 μM CaCl2 solution was added to the 96-well plate, and the plate was placed in a 37° C. incubator to pre-warm.d) 100 μL of the reaction mixture was added to the 96-well plate, and the plate was centrifuged at 1000 rpm for 10 seconds.e) The plate was read continuously for 10 min with a 30 seconds interval on a SpectraMax 340PC, with an instrument temperature of 37° C. and a wavelength of 360 nm. |
| Affinity data for this assay | |
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