| Assay Method Information | |
| | STING-Binding Test |
| Description: | Test compound (2 μL), Streptavidin-Terbium (4 μL, Cisbio), Fluorescein-labeled 2′,3′-cGAMP (c[G(2′,5′)p-2′-Fluo-AHC-A(3′,5′)p]) (Biolog, Germany) and biotinylated STING protein (2 μL, wild-type, WT) were mixed using assay buffer (Dulbecco's Phosphate-Buffered Saline (Wako Pure Chemical Industries, Ltd.) containing 0.01% bovine serum albumin free of fatty acid (Wako Pure Chemical Industries, Ltd.)), and the mixture was left to stand at room temperature for 3 hr (The final concentration. Streptavidin-Terbium; diluted by 1/1000, FITC-cGAMP; 1 μM, biotinylated STING protein; 100 nM). The time-resolved fluorescence resonance energy transfer (TR-FRET) was measured at the wavelength of 520 nm and 486 nm by EnVision (PerkinElmer, Waltham, MA, US). The inhibition rate of the binding of wild-type STING protein and 2′,3′-cGAMP of the test compound was calculated using the ratio of the count at 520 nm divided by the count at 486 nm. |
| Affinity data for this assay | |
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