ANKRD1 Colorimetric Cell-Based ELISA

From Assay Genie, the ANKRD1 Colorimetric Cell-Based ELISA is a high-throughput, lysate-free assay for measuring the relative amounts of ANKRD1 protein expression in cultured cells. The kit uses an indirect ELISA format: ANKRD1 is captured by target-specific primary antibodies and detected using HRP-conjugated secondary antibodies, with colorimetric readout at 450 nm. It is suitable for screening the effects of treatments, inhibitors such as siRNA or chemicals, and activators on ANKRD1 levels. The dynamic range is greater than 5000 cells, and the kit is reactive with Human, Mouse, and Rat samples, and is intended for adherent cells. A monoclonal antibody specific for human GAPDH is included as an internal positive control, and Crystal Violet whole-cell staining is used to determine cell density for normalization. The assay is run on a 96-well microplate, with storage at 4°C and a stated stability of 6 months.

ANKRD1 (ankyrin repeat domain-containing protein 1) is encoded on human chromosome 10q23.31. The protein is localized to the nucleus of endothelial cells and is induced by IL-1 and TNF-alpha stimulation. According to the cited references, it may act as a transcription factor involved in endothelial cell activation and in the negative regulation of cardiac gene expression. Defects in ANKRD1 have been associated with total anomalous pulmonary venous return (TAPVR), a rare congenital heart disease in which the pulmonary veins fail to connect to the left atrium during cardiac development.

The kit is supplied with a 96-well clear-bottom microplate suitable for cell culture, 10X TBS, quenching and blocking buffers, 15X wash buffer, primary and secondary antibody diluents, anti-phospho target antibody, anti-target antibody, anti-GAPDH antibody, HRP-conjugated anti-rabbit and anti-mouse IgG antibodies, SDS solution, stop solution, ready-to-use substrate, Crystal Violet solution, and adhesive plate seals.