GK2 Colorimetric Cell-Based ELISA

From Assay Genie, this cell-based ELISA kit is configured for the qualitative detection of Glycerol kinase 2 (GK2) in cultured human cells, using a colorimetric readout measured at 450 nm. The assay runs on a 96-well microplate format and is described as lysate-free and high throughput, suitable for measuring relative GK2 expression in adherent cells as well as screening the effects of treatments, inhibitors (such as siRNA or chemicals), or activators.

Detection uses an indirect ELISA format: GK2 is captured by target-specific primary antibodies and bound by HRP-conjugated secondary antibodies, which catalyze a colorimetric reaction upon substrate addition. Because the readout is qualitative, two normalization strategies are included — a monoclonal anti-GAPDH antibody serves as an internal positive control for target absorbance values, and a Crystal Violet whole-cell staining step can be used to determine cell density and correct for plating differences.

Manufacturer specifications indicate a dynamic range of "> 5000 Cells" and storage at 4°C with a stated 6-month stability. The kit is reactive with human samples and is intended for metabolism research. Reagents provided include 100x Anti-Phospho Target Antibody, 100x Anti-Target Antibody, Anti-GAPDH Antibody, HRP-Conjugated Anti-Rabbit IgG Antibody, HRP-Conjugated Anti-Mouse IgG Antibody, blocking and wash buffers, substrate, stop solution, and adhesive plate seals. A microplate reader capable of measuring absorbance at 450 nm is required but not supplied.