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. Author manuscript; available in PMC: 2016 Jul 1.
Published in final edited form as: J Biomol Screen. 2015 Mar 9;20(6):801–809. doi: 10.1177/1087057115575150

Figure 1.

Figure 1

Development of the enzyme-coupled SAMHD1 assay. (A) A standard curve for the detection of inorganic phosphate by malachite green shows a linear response up to 12 nmol of Pi (90 min incubation), which is the maximal amount of phosphate that could be generated in the coupled reaction of SAMHD1 and PPase in the high-throughput screen. (B) The rate of phosphate production in the SAMHD1 reaction is linear to 80% dGTP substrate conversion with only trace background activity from PPase. (C) Whole-plate replicates of the high-throughput screening reactions in the presence and absence of SAMHD1 showed small well-to-well variability and a large signal change for the SAMHD1 reaction (see Table 1). (D) Histogram plots the positive and negative control data in (C) showed normal distributions.