| Solution Information | help | |
| Enzyme: | Beta-galactosidase | |
| inhibitor: | BDBM48026 | |
| 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: Hams F-12 supplemented with 2.5% hiFBS, 1X Pen/Strep/Glu, 3) DiscoverX b-Arrestin Detection Reagents: Galacton Star, Emerald II, and Cell Assay Buffer Dose-Response Protocol: I. Enzyme addition 1. Using Biotek dispenser, dispense 6.0 ul of assay media to columns 1-4 and 6.0 uL of galactosidase enyme in assay buffer to columns 5-48 of a Corning, white 1536-well plate. Final concentration of enzyme is 0.05 units/ml. This concentration gave ~5% enzyme turnover in 15min. II. Compound Addition: 2. Using LabCyte Echo 555, transfer 40 nL of DMSO to positive and negative control wells in columns 1 - 4 and 45-48, respectively. Using a dose response protocol, transfer compounds from 10mM and 0.312 mM Echo qualified plates into assay plate columns 5 - 45. (Final concentrations range 66 uM to 0.128 uM, 10 doses, with 0.66% DMSO.) 3. Centrifuge plates at 500 rpm for 1 min. II. Set up of enzyme assay: 4. | |
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