S6 Ribosomal Protein (Phospho-Ser235)Colorimetric Cell-Based ELISA Kit

Manufactured by Assay Genie, this Colorimetric Cell-Based ELISA Kit is designed for the accurate detection of phosphorylated serine 235 levels of S6 Ribosomal Protein in cellular extracts. The kit offers a convenient, lysate-free, high-throughput and sensitive workflow for detecting S6 Ribosomal Protein phosphorylation and expression profiles in cells, and can be used to measure the relative amounts of phosphorylated S6 Ribosomal Protein in cultured cells or to screen the effects of various treatments, inhibitors (e.g., siRNA or chemicals) or activators on S6 Ribosomal Protein phosphorylation.

Reactive with Human, Mouse and Rat samples, the assay has a dynamic range of more than 5000 cells and uses an indirect ELISA format in which the target is captured by S6 Ribosomal Protein (Phospho-Ser235)-specific primary antibodies and detected by HRP-conjugated secondary antibodies that catalyse a colorimetric reaction read at 450 nm. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining step may be used to determine cell density. The target gene is RPS6 (Gene ID: 6194; UniProt ID: P62753); S6 Ribosomal Protein may play an important role in controlling cell growth and proliferation through the selective translation of particular classes of mRNA. The procedure is intended for adherent cells, beginning with seeding 200 µL of 20,000 adherent cells per well of a 96-well plate, fixation with formaldehyde, overnight primary antibody incubation at 4 °C, and a final absorbance readout at 450 nm after substrate addition.