GRK3 Colorimetric Cell-Based ELISA

From Assay Genie, this Cell-Based ELISA kit is designed for the precise measurement of G Protein-Coupled Receptor Kinase 3 (GRK3) levels in cell lysates and tissue homogenates. The assay uses a colorimetric detection method read at 450 nm in a 96-Well Microplate format, with reactivity validated for Human, Mouse, and Rat samples and adherent cells supported as the sample type.

The kit uses an indirect ELISA format in which GRK3 is captured by GRK3-specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody to catalyze a colorimetric reaction upon substrate addition. It is described as a convenient, lysate-free, high throughput and sensitive assay that detects the GRK3 protein expression profile in cells, and is suited to measuring the relative amounts of GRK3 in cultured cells as well as screening for the effects of treatments, inhibitors (such as siRNA or chemicals), or activators. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalising the target absorbance values, and the Crystal Violet whole-cell staining method may be used to determine cell density, by which the plating difference can be adjusted.

GRK3, also referred to as ADRBK2 or Beta-adrenergic receptor kinase 2, is a key enzyme involved in the regulation of G protein-coupled receptor signalling, with dysregulation linked to cancer, cardiovascular disorders, and neurological disorders. The protein has a reported molecular weight of 79,710 Da and is associated with protein amino acid phosphorylation, receptor internalisation, and signal transduction. The kit has a dynamic range of > 5000 Cells, with storage at 4 °C for 6 months.