| Assay Method Information | |
| | TRPV1 Inhibition Assay |
| Description: | Assay buffer preparation: 1 HBSS, 2 mM HEPES, 0.1% BSA plus 2.5 mM freshly prepared probenecid (Invitrogen, #P36400). 25 μl/well assay buffer in 384 well clear plate (cat #782281, Greiner Bio-one) was dispensed with buffer distributor (Multidrop ComB1 from Thermo Scientific). The compounds were in 384 Echo plate (cat #LPL0200, Labcyte) and the starting concentration of compound was 10 mM, then 1 to 3 serial dilution in 100% DMSO, 8 ul/well). 125 nl compounds was transferred to the 384 well plate containing 25 μl/well buffer with Echo 555 Liquid Handler (Labcyte), such that the compound concentration was 5 fold of final concentration. The plate was shook slowly at 40 rpm/min for 10 min to mix. 10 μl of 5 fold compound in the buffer was transferred to the cell plate (containing 20 μl cells and 20 μl dye) using Vertical Pipetting Station 384ST (Agilent Technologies). Six fold of final concentration of NADA (N-arachidonyl dopamine, cat #A8848, Sigma) in the assay buffer was prepared and 30 μl/well was distributed in 384 well clear plate. |
| Affinity data for this assay | |
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