CKI-alpha Colorimetric Cell-Based ELISA

From Assay Genie, this Cell-Based ELISA kit is designed for the precise quantification of CKI-alpha levels in cell lysates and tissue extracts using a colorimetric detection method read at 450 nm. The kit offers a lysate-free, high-throughput workflow in a 96-well microplate format and shows reactivity with samples of human, mouse, and rat origin, making it appropriate for adherent cell culture work.

Detection is achieved through an indirect ELISA format in which CKI-alpha-specific primary antibodies are bound by HRP-conjugated secondary antibodies that catalyze a colorimetric reaction upon substrate addition. An anti-GAPDH antibody is included as an internal positive control for normalizing target absorbance values, and a Crystal Violet whole-cell staining method can be used to determine cell density and adjust for plating differences. Storage conditions are 4°C for 6 months, and the dynamic range is greater than 5000 cells.

CKI-alpha (Casein kinase I isoform alpha, gene symbol CSNK1A1, UniProt P48729) is an ubiquitous protein kinase of the CK1 family that plays a critical role in regulating circadian rhythms and cell cycle progression and participates in Wnt signaling by phosphorylating beta-catenin, marking it for proteasomal degradation. Dysregulation of CKI-alpha has been linked to diseases such as cancer, neurological disorders, and metabolic conditions.