ChemiSCREEN Recombinant Human β2 Adrenoceptor

FUNCTION: SwissProt: P35626 # Specifically phosphorylates the agonist-occupied form of the beta-adrenergic and closely related receptors.
SIZE: 688 amino acids; 79710 Da
SIMILARITY: SwissProt: P35626 ## Belongs to the protein kinase superfamily. AGC Ser/Thr protein kinase family. GPRK subfamily. & Contains 1 AGC-kinase C-terminal domain. & Contains 1 PH domain. & Contains 1 protein kinase domain. & Contains 1 RGS domain.

Radioligand binding assay, and GTPγS binding.

One package contains enough membranes for at least 200 assays (units), where an unit is the amount of membrane that will yield greater than 40-fold signal:background with ¹²⁵I-labeled (-)Iodocyanopindolol at 0.5 nM. Liquid in packaging buffer: 50 mM Tris pH 7.4, 10% glycerol and 1% BSA with no preservatives. Packaging method: Membranes protein were adjusted to the indicated concentration in packaging buffer, rapidly frozen, and stored at -80oC.

Maintain frozen at -70°C for up to 2 years. Do not freeze and thaw.

Membranes are mixed with radioactive ligand and unlabeled competitor (see Figures 1 and 2 for concentrations tested) in binding buffer in a nonbinding 96-well plate, and incubated for 1-2 h. Prior to filtration, a GF/C 96-well filter plate is coated with 0.33% polyethyleneimine for 30 min, then washed with 50mM HEPES, pH 7.4. Binding reaction is transferred to the filter plate, and washed 3 times (1 mL per well per wash) with Wash Buffer. The plate is dried and counted.

Binding buffer: 50 mM Hepes, pH 7.4, 5 mM MgCl₂, 1 mM CaCl₂, filtered and stored at 4°C
Radioligand: [¹²⁵I]-(-) Iodocyanopindolol (Perkin Elmer # NEX189)
Wash Buffer: 50 mM Hepes, pH 7.4, 500 mM NaCl, filtered and stored at 4°C.

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