SEPARASE Colorimetric Cell-Based ELISA Kit

Designed for the accurate measurement of separase levels in cell lysates, this Cell-Based ELISA Kit from Assay Genie uses a colorimetric detection method read at 450 nm. It provides a convenient, lysate-free, high-throughput and sensitive assay that can detect SEPARASE protein expression profile in cells, enabling measurement of relative SEPARASE amounts in cultured cells as well as screening for the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on SEPARASE. The assay is reactive with human and mouse samples, has a dynamic range of > 5000 cells, and is intended for use with adherent cells.

In the indirect ELISA format, SEPARASE is captured by SEPARASE-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, allowing the conjugated HRP to catalyze a colorimetric reaction upon substrate addition. 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 method is provided to determine cell density so that absorbance values can be normalized to cell amounts. The kit is formatted on a 96-well microplate and is stored at 4 °C with a stated stability of 6 months.

Separase is a caspase-like protease that plays a central role in chromosome segregation by cleaving the SCC1/RAD21 subunit of the cohesin complex at the onset of anaphase, and is encoded by the ESPL1 gene (Gene ID: 9700, UniProt ID: Q14674). The kit is categorized within the research area of Epigenetics and Nuclear Signaling.