PFKFB2 (Phospho-Ser483)Colorimetric Cell-Based ELISA Kit

A cell-based ELISA from Assay Genie designed for the accurate detection of phosphorylated PFKFB2 protein levels in cell lysates. The assay is reactive with human, mouse, and rat samples, employs a colorimetric method measured at 450 nm, has a dynamic range of > 5000 Cells, and is intended for use with adherent cells.

The assay is lysate-free, high-throughput, and sensitive, enabling detection of PFKFB2 protein phosphorylation and expression profiles in cultured cells through an indirect ELISA format. Phosphorylated PFKFB2 at Ser483 is captured by phospho-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody to catalyze a colorimetric reaction upon substrate addition. The kit can be used to measure relative amounts of phosphorylated PFKFB2 in cultured cells and to screen for the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on PFKFB2 phosphorylation. A monoclonal antibody specific for human GAPDH is included as an internal positive control, and a Crystal Violet whole-cell staining method can be used to determine cell density, with absorbance values normalized to cell amounts to adjust for plating differences.

PFKFB2 is a bifunctional enzyme that participates in both the synthesis and degradation of fructose-2,6-bisphosphate, a regulatory molecule controlling glycolysis in eukaryotes. The protein has a molecular weight of 54,406 Da and is encoded by the PFKFB2 gene (Gene ID: 5208, UniProt ID: O60825). The enzyme has been implicated in various diseases, including cancer and metabolic disorders, and monitoring its phosphorylation status at Ser483 can provide insights into its role in cellular processes and inform the development of targeted therapies and diagnostic tools.