| Solution Information | help | |
| Enzyme: | Beta-galactosidase | |
| inhibitor: | BDBM67390 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay materials: 1) Beta-Galactosidase from Sigma-Aldrich (Cat# G4155) 2) Assay Medium: Opti-MEM Medium supplemented with 1% hiFBS, 1X Pen/Strep/Glu, 125 ug/mL Hygromycin (1/2 recommended), 250 ug/mL Geneticin (1/2 recommended) 3) DiscoverX b-Arrestin Detection Reagents: Galacton Star, Emerald II, and Cell Assay Buffer Dose Response protocol: 1) Add 5.0 uL of b-gal diluted in assay media to 0.03 U/mL to all wells (0.02 U/mL final assay concentration) of a 1536-well plate with the exception of the positive control wells where 5.0 uL of assay media alone is added. 2) Spin plates at 500 rpm for 1 min. 3) Prepare Detection Reagent Solution from DiscoveRx (1 part Galacton Star: 5 parts Emerald II and 19 parts Cell Assay Buffer). 4) Using a Labcyte Echo, DMSO and test compounds are transferred to assay wells. DMSO only is transferred to positive and negative control wells, while varying volumes of test compounds are transferred to test compound wells to achieve the desired test concentrat | |
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