E2F1 (Phospho-Thr433)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA Kit is designed for the precise detection of E2F1 phosphorylation at threonine 433 in cells. The assay uses a colorimetric detection method read at 450 nm in a 96-well microplate format, is reactive with human and mouse samples, and has a dynamic range of > 5000 Cells. The kit provides a convenient, lysate-free workflow for measuring relative amounts of phosphorylated E2F1 in cultured cells and for screening the effects of various treatments, inhibitors, or activators on E2F1 phosphorylation.

E2F1 is a transcription factor of the E2F/DP family that regulates cell cycle progression and is a target of tumour suppressor proteins. Phosphorylation of E2F1 at threonine 433 has been linked to its function in cell proliferation and the DNA damage response, making it a relevant biomarker in cancer biology research. The assay uses an indirect ELISA format in which E2F1 (Phospho-Thr433) is captured by target-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody and catalyse 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. Following the colorimetric measurement of HRP activity, the Crystal Violet whole-cell staining method can be used to determine cell density, allowing absorbance values to be normalised to cell amounts and adjusted for plating differences.