SIAH1 Colorimetric Cell-Based ELISA

A cell-based ELISA kit from Assay Genie designed for the quantitative measurement of SIAH1 protein levels in cell lysates. The kit offers a lysate-free, high-throughput and sensitive indirect ELISA format in which SIAH1 is captured by target-specific primary antibodies and detected using HRP-conjugated secondary antibodies that catalyze a colorimetric reaction at 450 nm upon substrate addition. Reactivity is reported for Human, Mouse and Rat samples, and the assay supports a dynamic range of > 5000 cells. The stated sample type is adherent cells, and the kit is provided in a 96-well microplate format with a storage recommendation of 4 °C for 6 months.

The kit can be used to measure relative amounts of SIAH1 in cultured cells as well as to screen the effects that various treatments, inhibitors (such as siRNA or chemicals) or activators have on SIAH1. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalizing the target absorbance values. Following the colorimetric measurement of HRP activity via substrate addition, the Crystal Violet whole-cell staining method may be used to determine cell density, with results analysed by normalizing absorbance values to cell amounts so that plating differences can be adjusted.

SIAH1 is a key regulator of the ubiquitin-proteasome pathway, functioning as an E3 ubiquitin-protein ligase that mediates ubiquitination and subsequent proteasomal degradation of target proteins. It has a critical role in processes such as cell cycle progression, apoptosis and cellular signaling, and its dysregulation has been implicated in cancer, neurodegenerative disorders and inflammatory conditions, highlighting its relevance as a potential therapeutic target and biomarker for disease progression.