| Assay Method Information | |
| | ACLY Enzyme Activity Assay |
| Description: | Experimental method: ADP Glo luminescence method was used for determination. It reflects the activity of ACLY enzyme by quantitatively detecting the amount of ADP, and the enzymatic reaction catalyzed by ACLY is proportional to the amount of ADP detected by the luminescent signal. Firstly, the compound is diluted with 10% DMSO, and then 11 of compound dilution is added to a 5 μl reaction system, so that the final reaction system has a DMSO content of 2%. The enzymatic reaction catalyzed by ACLY was carried out at 37° C. for 30 minutes. 5 μl reaction mixture contained the following components: 40 mM Tris, pH 8.0, 10 mM MgCl2, 5 mM DTT, ATP, CoA, sodium citrate, and ACLY. After the enzymatic reaction was complete, 2.5 μl of ADP Glo reagent was added to each reaction system and incubated at room temperature for 1 hour. Afterwards, 5 μl of kinase detection reagent was added and incubated at room temperature for 30 minutes. The luminous signal was detected using Envision (PerkinElmer, USA). |
| Affinity data for this assay | |
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