AKAP13 Colorimetric Cell-Based ELISA

From Assay Genie, this cell-based ELISA kit enables the qualitative detection of AKAP13 (A-kinase anchor protein 13) protein expression in cultured cells. It is designed for the precise measurement of AKAP13 levels in cell lysates and tissue samples, and can be used to assess relative AKAP13 expression in cultured cells as well as to screen the effects of treatments, inhibitors such as siRNA or chemicals, or activators. AKAP13 anchors cAMP-dependent protein kinase (PKA), couples G alpha-13 and Rho, and is involved in protein-protein interactions and the localisation of signalling molecules, making it a relevant target for studies of cell signalling and disease pathways.

The assay uses an indirect ELISA format in which AKAP13-specific primary antibodies capture the target, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. On substrate addition, the HRP enzyme catalyses a colour change read at 450 nm. The kit is reactive with human samples, provided in a 96-well microplate format, with a stated dynamic range of > 5000 Cells and recommended storage at 4 °C for 6 months. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance, and Crystal Violet whole-cell staining can be used to determine cell density and adjust for plating differences.

Sample type is adherent cells, and the workflow is lysate-free and high-throughput, with cells fixed directly in the plate, quenched, blocked, and incubated with primary antibody overnight at 4 °C before detection.