SEPARASE (Phospho-Ser801)Colorimetric Cell-Based ELISA Kit

This Cell-Based ELISA Kit from Assay Genie is designed for the detection of phospho-serine 801 levels of SEPARASE in cell lysates and cell culture supernatants, with a colorimetric detection method read at 450 nm. The kit shows human reactivity, has a dynamic range of > 5000 cells, and is supplied as 2 x 96-Well Microplates for adherent cell samples.

Separase is a caspase-like protease encoded by the ESPL1 gene (Gene ID: 9700, UniProt ID: Q14674) that cleaves the SCC1/RAD21 subunit of the cohesin complex at the onset of anaphase, initiating the final separation of sister chromatids. Phosphorylation at Ser801 is a critical post-translational modification that regulates Separase activity, making it a relevant marker for studying cell cycle progression and genomic stability. The product is classified under the Phospho Specific and Epigenetics and Nuclear Signaling research areas.

The assay uses an indirect ELISA format in which SEPARASE (Phospho-Ser801) is captured by target-specific primary antibodies, then bound by HRP-conjugated secondary antibodies that catalyse a colorimetric reaction upon substrate addition. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalising absorbance values, and Crystal Violet whole-cell staining can be used to determine cell density for further normalisation. The procedure involves seeding adherent cells at 20,000 cells per well in 200 µL of culture medium, overnight incubation at 4 °C with the primary antibody, 1.5 hours with the secondary antibody at room temperature, and a 30-minute substrate development step before reading the optical density at 450 nm.