IPMK Colorimetric Cell-Based ELISA

Assay Genie provides a cell-based colorimetric ELISA for the qualitative detection of inositol polyphosphate multikinase (IPMK) in cultured cells, including adherent cell samples. The assay is read at 450 nm, has a dynamic range of > 5000 cells, and is reactive with human, mouse, and rat samples, supporting metabolism research. The kit is described as convenient, lysate-free, high throughput and sensitive, and is intended for measuring IPMK protein expression profiles in cells and for screening the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on IPMK expression.

Detection follows an indirect ELISA format: IPMK is captured by IPMK-specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. Upon substrate addition, the conjugated HRP enzyme catalyses a colorimetric reaction. Because the assay is qualitative, two normalisation methods are included. First, a monoclonal antibody specific for human GAPDH serves as an internal positive control for normalising target absorbance values. Second, a Crystal Violet whole-cell staining step allows normalisation of absorbance values to cell density, adjusting for plating differences between wells.

IPMK (inositol polyphosphate multikinase) is an inositol phosphate kinase with broad substrate specificity, classified under EC 2.7.1.151 with a molecular weight of 47,222 Da. The human ortholog is located at chromosomal position 10q21.1 (Gene ID: 253430; UniProt: Q8NFU5). The encoded protein has 3-kinase, 5-kinase and 6-kinase activities on phosphorylated inositol substrates and participates in inositol phosphate biosynthesis and metabolic processes. According to the manufacturer, dysregulation of IPMK has been linked to cancer, diabetes, and neurological disorders, making it relevant to research on cellular signalling, growth, proliferation, and metabolism. The kit is stored at 4 °C with a 6-month shelf life.