PBK/TOPK (Phospho-Thr9)Colorimetric Cell-Based ELISA Kit

A Cell-Based ELISA from Assay Genie, this kit is designed for the detection of phosphorylated Thr9 on the PBK/TOPK protein in cell lysates. The Colorimetric assay is read at 450 nm and is reactive with human samples, with a stated dynamic range of > 5000 Cells and supplied in a 2 x 96-Well Microplate format.

The kit provides a convenient, lysate-free, high throughput and sensitive method for detecting PBK/TOPK protein phosphorylation and expression profile in cells. It can be used to measure relative amounts of phosphorylated PBK/TOPK in cultured cells and to screen the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on PBK/TOPK phosphorylation. Detection is achieved via an indirect ELISA format: PBK/TOPK (Phospho-Thr9) is captured by target-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, allowing the conjugated HRP to catalyse a colourimetric reaction upon substrate addition.

A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control for normalising target absorbance values. Following the colourimetric measurement of HRP activity via substrate addition, Crystal Violet whole-cell staining may be used to determine cell density; the absorbance values can then be normalised to cell amounts to adjust for plating differences. PBK (Gene ID: 55872, UniProt ID: Q96KB5, chromosomal location 8p21.2) is a serine/threonine kinase implicated in cell proliferation, migration and survival, with phosphorylation at Thr9 linked to cellular processes and diseases including cancer and neurodegenerative disorders.