PTTG1 Colorimetric Cell-Based ELISA

This Assay Genie kit is a cell-based ELISA for the qualitative measurement of human PTTG1 (Pituitary Tumor-Transforming Gene 1) in adherent cells. The assay uses an indirect ELISA format in which PTTG1 is captured by target-specific primary antibodies and detected with HRP-conjugated secondary antibodies that catalyze a colorimetric reaction upon substrate addition, read at 450 nm. The dynamic range is > 5000 cells.

The protocol is lysate-free and supports high-throughput screening of the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on PTTG1 expression in cultured cells. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalizing target absorbance values. After the colorimetric read, a Crystal Violet whole-cell staining step can be used to determine cell density, allowing absorbance values to be normalized to cell number and plating differences to be adjusted. The kit is intended for cell cycle research and is stored at 4 °C for up to 6 months.

PTTG1 encodes securin, a regulatory protein involved in chromosome stability, the p53/TP53 pathway, and DNA repair. During mitosis it inhibits Separase/ESPL1 to prevent premature cohesin cleavage and is itself a substrate of the anaphase-promoting complex. It is of interest as a biomarker in cancer research and drug development.