Interested in assay kits to assess DDX1 & DDX41?
Activity was measured using the Transcreener® ADP2 FP Assay using 12 nM human DDX1 (BellBrook Labs #2305). SCH-202676 was serially diluted and incubated with enzyme for 30 minutes. The following was added to start the reaction: 15 µM ATP, 0.5 mg/mL yeast RNA, 50 mM TRIS pH 7.5, 2 mM MgCl2, 0.01% Triton X-100; 10 μL Enzyme Reaction incubated @ 30°C for 60 minutes in a Corning® low-volume 384-well plate (#4514). 10 μL of the following detect mix was added: 16 μg/ mL ADP2 Ab, 4 nM ADP2 Tracer, 1X Stop and Detect Buffer B. 20 μL Complete Reaction was incubated for 60 minutes and read on a CLARIOstarPlus instrument. Fluorescent Polarization values were converted to ADP formed with a 10 μM ADP Standard Curve and was plotted using Prism (GraphPad). An IC50 of 0.86 µM was obtained.
Activity was measured using the Transcreener® ADP2 FP Assay using 100 nM human DDX41 (BellBrook Labs #2315). SCH-202676 was serially diluted and incubated with enzyme for 30 minutes. The following was added to start the reaction: 15 µM ATP, 0.5 mg/mL yeast RNA, 50 mM TRIS pH 7.5, 2 mM MgCl2, 0.01% Triton X-100; 10 μL Enzyme Reaction incubated @ 30°C for 60 minutes in a Corning® low-volume 384-well plate (#4514). 10 μL of the following detect mix was added: 16 μg/ mL ADP2 Ab, 4 nM ADP2 Tracer, 1X Stop and Detect Buffer B. 20 μL Complete Reaction was incubated for 60 minutes and read on a CLARIOstarPlus instrument. Fluorescent Polarization values were converted to ADP formed with a 10 μM ADP Standard Curve and was plotted using Prism (GraphPad). An IC50 of 1.78 µM was obtained.
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BellBrook Labs
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Enzolution® TREX1 Assay System
