AKAP14 Colorimetric Cell-Based ELISA

The Assay Genie AKAP14 Colorimetric Cell-Based ELISA Kit is a cell-based immunoassay for the detection of AKAP14 protein levels in cell lysates and cell culture supernatants from adherent cells. The assay uses an indirect ELISA format, in which AKAP14 is captured by target-specific primary antibodies and detected via HRP-conjugated secondary antibodies; signal is generated by a colorimetric reaction read at 450 nm. A GAPDH-specific monoclonal antibody is included as an internal positive control, and Crystal Violet whole-cell staining is used to normalize absorbance values to cell density.

AKAP14 (A-kinase anchor protein 14) functions as a scaffold for protein kinase A (PKA) signaling complexes, binding type II regulatory subunits of PKA and anchoring the holoenzyme to discrete cellular locations. It is involved in processes such as cell proliferation, apoptosis, and gene expression regulation, and is associated with spermatogenesis. The protein is localised to the cytoplasm with a molecular weight of 10,187 Da, and 3 isoforms of the human protein are produced by alternative splicing. According to NCBI, the encoded protein anchors PKA in ciliary axonemes and may play a role in regulating ciliary beat frequency.

The kit can be used both to measure relative AKAP14 amounts in cultured cells and to screen for the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators. The dynamic range is reported as > 5000 cells, and the assay is supplied in a 96-well microplate format. It is intended for research applications including studies of AKAP14-related mechanisms in cancer, cardiovascular disease, and neurological disorders. The kit should be stored at 4°C with a stated stability of 6 months.