DGKI Colorimetric Cell-Based ELISA

This Cell-Based ELISA kit from Assay Genie is designed for the qualitative detection of DGKI protein expression in cultured cells. The assay uses a colorimetric detection method with measurement at 450 nm and an indirect ELISA format, in which DGKI-specific primary antibodies are captured while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. The HRP enzyme conjugated to the secondary antibody catalyzes a colorimetric reaction upon substrate addition. The kit is reactive with human and rat samples and is suitable for adherent cells.

The dynamic range of the assay is greater than 5000 cells. The kit is provided in a microplate format and is stored at 4°C with a stability of 6 months. It is a lysate-free assay that can be used to measure the relative amounts of DGKI in cultured cells and to screen for the effects of various treatments, inhibitors, or activators on DGKI expression.

For data normalization, 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, and results can be analysed by normalizing the absorbance values to cell amounts to adjust for plating differences. The target DGKI corresponds to Gene ID 9162 and UniProt ID O75912, with a molecular weight of 116,997 Da.