Clock Colorimetric Cell-Based ELISA

From Assay Genie, this In-Cell ELISA Kit provides a colorimetric Cell-Based assay for measuring the relative levels of the Clock protein in cultured cells, with detection read at 450 nm. The lysate-free, high-throughput format supports qualitative detection of Clock expression profiles and can be used to screen the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on Clock in adherent cell cultures, with reactivity demonstrated in Human, Mouse, and Rat samples.

The assay follows an indirect ELISA principle: Clock is captured by target-specific primary antibodies, and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, enabling a colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included as an internal positive control for normalizing target absorbance values, and a Crystal Violet whole-cell staining step is provided to determine cell density. Specified performance attributes include a Dynamic Range of > 5000 Cells, a Colorimetric detection method at 450 nm, and storage at 4°C for 6 months.

The CLOCK target is a basic helix-loop-helix transcription factor with intrinsic histone acetyltransferase activity that functions as a core component of the circadian oscillator. CLOCK forms heterodimers with ARNTL/2 to activate E-box-dependent transcription of PER1 and PER2, with feedback inhibition by PER and CRY proteins. Database references associated with the target include Gene ID 9575, UniProt ID O15516, OMIM 601851, and Unigene Hs.436975.