ANKK1 Colorimetric Cell-Based ELISA

Assay Genie's cell-based ELISA kit provides a colorimetric method for detecting ANKK1 protein in human cells, with absorbance read at 450 nm. The qualitative, indirect ELISA format enables measurement of relative ANKK1 levels in cultured cells and screening of how various treatments, inhibitors such as siRNA or chemicals, and activators affect ANKK1 expression, without requiring lysate preparation. ANKK1-specific primary antibodies capture the target, and HRP-conjugated secondary antibodies catalyse the colorimetric reaction upon substrate addition. ANKK1 belongs to the Ser/Thr protein kinase family and has been implicated in dopamine receptor signalling and in neuropsychiatric disorders such as schizophrenia and addiction.

Running on a 96-well microplate format, the assay is designed for adherent cells and has a dynamic range greater than 5000 cells. Storage is at 4 °C for 6 months. Two normalisation approaches are built in: a monoclonal antibody specific for human GAPDH acts as an internal positive control to normalise target absorbance, and a Crystal Violet whole-cell stain can be used to determine cell density so that absorbance values can be adjusted for plating differences.

Target background: ANKK1, also known as ankyrin repeat and protein kinase domain-containing protein 1 (synonyms include protein kinase PKK2, Sugen kinase 288, sgK288, and X-kinase), is encoded at chromosome 11q23.2 in close linkage to DRD2. The protein has a reported molecular weight of 84,632 Da, with database identifiers Gene ID 255239 and UniProt ID Q8NFD2.