ATPase Colorimetric Cell-Based ELISA

From Assay Genie, this Cell-Based ELISA kit is designed for the measurement of ATPase activity in cell lysates and tissue homogenates. The assay uses an indirect ELISA format in which ATPase is captured by target-specific primary antibodies and detected via HRP-conjugated secondary antibodies, producing a colorimetric readout at 450 nm after substrate addition. The kit is reactive with Human, Mouse, and Rat samples and is configured for a 96-well microplate format with a stated dynamic range of greater than 5000 cells.

A monoclonal antibody specific for human GAPDH is included as an internal positive control to help normalise target absorbance values, and a Crystal Violet whole-cell staining step may be used after the HRP colorimetric measurement to determine cell density. This allows absorbance values to be normalised to cell numbers, adjusting for plating differences. The kit is suited for adherent cells and is stored at 4°C for up to 6 months.

The target, ATPase, corresponds to the ATP1A1 gene product, sodium/potassium-transporting ATPase subunit alpha-1, which catalyses the hydrolysis of ATP coupled with the exchange of sodium and potassium ions across the plasma membrane. The kit falls within the Signal Transduction research area and can be applied to assess relative ATPase levels in cultured cells, as well as to screen the effects of treatments, inhibitors (such as siRNA or chemicals), or activators.