PAK1 Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA Kit is designed for the accurate detection of PAK1 (P21-activated kinase 1) in cell lysates and tissue homogenates, supporting research in cell signalling and protein kinase biology. The assay is reactive with Human, Mouse and Rat samples and uses a colorimetric detection method read at 450 nm, with a stated dynamic range of > 5000 Cells. The kit is supplied in a microplate format intended for use with adherent cells, with storage at 4°C for 6 months.

The workflow is lysate-free and offers a high-throughput, sensitive means of measuring relative PAK1 protein expression in cultured cells, as well as screening the effects of treatments, inhibitors (such as siRNA or chemicals) or activators on PAK1. Qualitative determination is achieved through an indirect ELISA format: PAK1-specific primary antibodies are bound by HRP-conjugated secondary antibodies, and the conjugated HRP enzyme catalyses a colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values, while a Crystal Violet whole-cell staining step allows absorbance values to be normalised to cell density, adjusting for plating differences.

PAK1 is a serine/threonine protein kinase of the STE20 family and a key effector of Rho GTPases. It regulates cytoskeleton reorganisation, the JNK MAPK pathway and nuclear signalling, and is implicated in processes including cell migration, proliferation and survival. Binding of Rac/cdc42 to the PAK1 p21-binding domain triggers autophosphorylation and conformational change, and the kinase is itself phosphorylated and activated by PDK.