ITCH Colorimetric Cell-Based ELISA Kit

From Assay Genie, this In-Cell ELISA kit enables qualitative detection of ITCH protein expression in cultured cells, including for screening the effects of treatments, inhibitors such as siRNA or chemicals, or activators on ITCH. The kit is suited to adherent cell samples and reacts with human and mouse material. Detection is colorimetric, read at 450 nm, using a 96-well microplate format with a stated dynamic range of > 5000 cells and recommended storage at 4°C for 6 months.

The assay uses an indirect ELISA format: ITCH is captured by ITCH-specific primary antibodies, and HRP-conjugated secondary antibodies then catalyze a colorimetric reaction upon substrate addition. Because the readout is qualitative, two normalization methods are provided — a monoclonal antibody specific for human GAPDH as an internal positive control, and a Crystal Violet whole-cell staining step to determine cell density, so that absorbance values can be normalized to cell amounts.

ITCH (NCBI Gene ID: 83737; UniProt: Q96J02) is described as an E3 ubiquitin-protein ligase with a reported molecular weight of 102 kDa, and the kit is positioned as a convenient, lysate-free, high throughput tool for studying this target.