Adrenergic Receptor beta2 Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA Kit is designed for the qualitative detection of Adrenergic Receptor beta2 (ADRB2) protein expression directly in cultured human cells, without a lysate preparation step. The kit uses an indirect ELISA format: the target protein is captured by Adrenergic Receptor beta2-specific primary antibodies and detected using HRP-conjugated secondary antibodies that catalyze a colorimetric reaction on substrate addition, with absorbance read at 450 nm. A reported dynamic range of "> 5000 Cells" is given for the assay, and a monoclonal anti-human GAPDH antibody is included to serve as an internal positive control. A Crystal Violet whole-cell staining step may be used to normalize target absorbance values to cell density, adjusting for differences in plating.

The assay is supplied in a 96-well microplate format and is suitable for measuring relative ADRB2 levels in adherent cells, as well as for screening the effects of various treatments, inhibitors (such as siRNA or chemicals) or activators on ADRB2. The procedure involves seeding cells, fixation with formaldehyde, quenching, blocking, primary antibody incubation (overnight at 4 °C, or 2 hours at room temperature for highly expressed targets), HRP-conjugated secondary antibody incubation, ready-to-use substrate addition and stop solution, with absorbance read immediately at 450 nm. Storage is at 4 °C with a stated stability of 6 months.

Reagents supplied include 10X TBS, quenching buffer, blocking buffer, 15X wash buffer, primary antibody diluent, 100X anti-phospho-target antibody, 100X anti-target antibody, anti-GAPDH antibody, HRP-conjugated anti-rabbit IgG, HRP-conjugated anti-mouse IgG, SDS solution, stop solution, ready-to-use substrate, crystal violet solution, 96-well cell culture clear-bottom microplates and adhesive plate seals. ADRB2 is a G-protein coupled receptor that mediates catecholamine-induced activation of adenylate cyclase and binds epinephrine with an approximately 30-fold greater affinity than norepinephrine. The source notes disease associations including obesity and susceptibility to asthma.