Assay Method Information

Assay Name:  heGAS Kinase-Glo Assay
Description:  Certain compounds of the present disclosure were tested for their h-cGAS inhibition activity using the methodology reported in Lama et al., “Development of human cGAS-specific small molecule inhibitors for repression of dsDNA-triggered interferon expression”, Nature Communications 10, Article number: 2261 (2019), with slight changes to some conditions as shown in Table B.TABLE BSummary of assay conditionsLama Presentet al. 2019 DisclosureEnzyme h-cGAS (nM) 100 40BufferTris-HCl pH 7.4 (mM) 20 20MgCl2 (mM) 5 10NaCl (mM) 150 25Tween ™-20 (%) 0.01 0.01ZnCl2 (μM) 1 1DTT (mM) 1 1DMSO (%) 0.5 5SubstratesATP (μM) 100 100GTP (μM) 100 100dsDNA (nM) 25 25AssayPlate (wells) 384 384Incubation length (h) 7 3Total volume (μL) 20 20Kinase-Glo Max (μL) 20 20
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