PAK3 (Phospho-Ser154)Colorimetric Cell-Based ELISA Kit

Designed for the precise measurement of phosphorylated PAK3 at Ser154 in cultured cells, this Colorimetric Cell-Based ELISA Kit from Assay Genie provides a convenient, lysate-free, high-throughput assay platform. The kit uses an indirect ELISA format in which PAK3 (Phospho-Ser154) is captured by target-specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, catalysing a colorimetric reaction at 450 nm upon substrate addition.

The assay is reactive with human, mouse, and rat samples and has a stated dynamic range of > 5000 cells. It is intended for adherent cell cultures and supports screening for the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on PAK3 phosphorylation. Two normalisation approaches are included: a monoclonal anti-GAPDH antibody serves as an internal positive control for normalising target absorbance, and a Crystal Violet whole-cell staining step is used to determine cell density so that absorbance values can be adjusted for plating differences.

PAK3 is a serine/threonine protein kinase of the STE20 family and a key regulator of cytoskeletal dynamics, cell migration, proliferation, and differentiation. Phosphorylation at Ser154 is reported to modulate its activity and influence downstream signalling pathways involved in cell growth and development, with implications in cancer, neurodegeneration, and developmental disorders. Missense and truncation mutations in PAK3 are linked to non-syndromic X-linked mental retardation type 30 (MRX30). Database identifiers for the target include Gene ID 5063, UniProt O75914, OMIM 300142/300558, and Unigene Hs.656789.