Dyskerin Colorimetric Cell-Based ELISA

Assay Genie provides a Cell-Based (In-Cell) ELISA Kit for the quantitative detection of dyskerin levels in cell lysates or tissue samples. Designed for adherent cells as a lysate-free, high-throughput assay, it measures relative dyskerin protein expression in cultured cells and supports screening of the effects that various treatments, siRNA, chemicals, or activators have on dyskerin.

Detection uses an indirect ELISA format, in which dyskerin is captured by dyskerin-specific primary antibodies and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, catalysing a colorimetric reaction upon substrate addition that is read at 450 nm. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalising target absorbance values, and the Crystal Violet whole-cell staining method may be used to determine cell density so that absorbance values can be normalised to cell amount, adjusting for plating differences. The kit shows reactivity with human, mouse and rat samples, with a dynamic range of greater than 5000 cells.

Dyskerin is encoded by the DKC1 gene (Gene ID: 1736, UniProt ID: O60832) and plays an active role in ribosome biogenesis and telomere maintenance, acting as a probable catalytic subunit of the H/ACA small nucleolar ribonucleoprotein complex that catalyses pseudouridylation of rRNA. The kit is configured in a 96-well microplate format, with storage at 4°C for up to 6 months.