| Solution Information | help | |
| Enzyme: | Beta-adrenergic receptor kinase 1 | |
| inhibitor: | BDBM79508 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | GRK2 protein is biotinylated using biotinamidohexanoic acid N-hydroxysuccinimide ester(Sigma). The RNA aptamer is fluorescently labeled on the 3'end with carboxyfluorescein (synthesized and labeled byIDT). Streptavidin-coated beads (Spherotech) are incubated with biotinylated GRK2 (bGRK2) at a final concentration of 2 nM for 30 minutes. The BioTek Microflow liquid dispenser is used to dispense 4 microL of assay buffer to all but column 1 of a 384-well assay plate. The positive (blocked) control containing 50X unlabeled RNA aptamer in assay buffer is dispensed to column 1 by a Microflow liquid dispenser (BiotTek, USA). Compounds in a dose response range of concentrations (33 microM to 5 nanoM in-well concentration) are transferred to assay wells via 100 nanoL pintool transfer on the Biomek FX liquid dispenser (Beckman Coulter, USA. A total of 3 microL of bead suspension is dispensed into assay wells using the Nanoquot liquid dispenser (BioTek, USA). Plates are incubated at RT for 3 | |
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