| Assay Method Information | |
| | MAT2A Biochemical Assay |
| Description: | Enzyme Reaction(1). Prepared 1×Assay buffer.(2). Preparation of compound concentration gradient: the compounds test condition were 1 uM start, 3-fold dilution, 10 doses, singlet or duplicate. 100× concentration compounds were prepared in 384-well plate. Then used Echo 550 to transfer 250 nl to a 384-reaction plate for later use. Added 250 nl of 100% DMSO to the negative and positive control wells.(3). Prepared 1.67× final concentration enzyme solution with 1×Assay buffer.(4). Added 15 μl of 1.67× Enzyme solution to the compound wells and positive control wells; added 15 μl of 1 Assay buffer to the negative control wells.(5). Centrifuged at 1000 rpm for 30 seconds and incubate for 15 minutes.(6). Prepared 2.5× final concentration Substrate mix solution with 1×Assay buffer.(7). Added 10 μl of 2.5× final concentration Substrate mix solution to start the reaction.(8). Centrifuged at 1000 rpm for 30 seconds and incubate for 150 minutes.(9). Added 50 μl Biomol Green to stop the reaction, centrifuge at 1000 rpm for 30 seconds and incubate for 15 minutes. read O.D.620, process data. |
| Affinity data for this assay | |
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