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. Author manuscript; available in PMC: 2016 Mar 23.
Published in final edited form as: J Biomol Screen. 2012 Dec 4;18(6):670–677. doi: 10.1177/1087057112469406

Figure 1. Assay miniaturization and validation.

Figure 1

Concentration responses for (a) relaxin and (b) forskolin in HEK293-RXFP1 cells. Assays were performed in 384-well and 1,536-well formats using the HTRF cAMP assay kit. 100% activity was normalized as 57.7 μM forskolin stimulation and 0% activity was normalized as 0.58% DMSO controls. (c) HTRF signal from 0.57% DMSO, 28.75 nM relaxin and 57.5 μM forskolin treatments in HEK293-RXFP1 cells. Mean and standard deviations were calculated from 12 treatment wells in 384-well assay format. (d) Scatter plot of the ratiometric signal from a 1,536-well DMSO plate to establish assay performance parameters. Column 1 was treated with a relaxin titration (24.9 pM – 1.47 μM), columns 2 and 3 received DMSO, and column 4 received 125 nM relaxin.