ALDOA Colorimetric Cell-Based ELISA

Assay Genie's cell-based ELISA kit is designed for the accurate quantification of ALDOA (Aldolase A) levels, providing researchers with a high-throughput, lysate-free assay. The kit measures relative ALDOA protein expression in cultured cells and is suitable for screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators. Detection is by a colorimetric method at 450 nm in a 96-well microplate format, with reactivity against human, mouse, and rat samples and a dynamic range of greater than 5000 cells.

The assay uses an indirect ELISA format: ALDOA is captured by ALDOA-specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. The HRP enzyme catalyses a colorimetric reaction upon substrate addition. Because the assay is qualitative, normalization is required. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control for normalizing target absorbance values. Following colorimetric measurement of HRP activity, the Crystal Violet whole-cell staining method may be used to determine cell density, and absorbance values can be normalized to cell amounts to adjust for plating differences.

ALDOA is a key glycolytic enzyme catalysing the reversible conversion of fructose-1,6-bisphosphate to glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. Dysregulation of ALDOA has been associated with cancer, metabolic disorders, and neurodegenerative diseases, making this kit a useful tool for researchers investigating the role of ALDOA in health and disease. The kit is stored at 4°C with a stated stability of 6 months.