Retinoblastoma (Phospho-Ser780)Colorimetric Cell-Based ELISA Kit

From Assay Genie, this Cell-Based ELISA Kit is designed for the qualitative determination of phosphorylated Retinoblastoma protein at serine 780 in cultured cells. The assay uses an indirect ELISA format in which target-specific primary antibodies capture the phospho-protein and HRP-conjugated secondary antibodies catalyze a colorimetric reaction upon substrate addition, read at 450 nm. The kit is described as convenient, lysate-free, high throughput and sensitive, with a stated dynamic range of > 5000 Cells and reactivity against human, mouse and rat samples. It is intended for use with adherent cells.

Two normalization strategies are built into the workflow. A monoclonal antibody specific for human GAPDH is included as an internal positive control to normalize target absorbance values, and a Crystal Violet whole-cell staining step can be used to determine cell density, allowing absorbance values to be adjusted for plating differences. The kit can be applied to measuring the relative amounts of phosphorylated Retinoblastoma in cultured cells and to screening the effects of various treatments, inhibitors (such as siRNA or chemicals) or activators on Retinoblastoma phosphorylation. The Retinoblastoma protein, encoded by the RB1 gene located at chromosomal position 13q14.2, is a tumor suppressor that regulates cell proliferation through the G1-phase restriction point, and phosphorylation at serine 780 is described as a key event in cell cycle control relevant to research into cancers including retinoblastoma.