MCM4 Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit is designed for the detection of MCM4 protein levels in cell samples using a colorimetric readout at 450 nm. The kit is reactive with both human and mouse samples and has a dynamic range of greater than 5000 cells. It is supplied in a microplate format and is intended to be stored at 4°C with a stated stability of 6 months. The specified sample type is adherent cells.

The assay is lysate-free and high throughput, allowing measurement of relative MCM4 amounts in cultured cells and screening of the effects that various treatments, inhibitors (such as siRNA or chemicals), or activators have on MCM4. Qualitative determination of MCM4 is achieved by an indirect ELISA format, in which MCM4 is captured by specific primary antibodies and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody; the conjugated HRP then catalyses a colorimetric reaction upon substrate addition. A monoclonal antibody specific for human GAPDH is included to serve as an internal positive control in normalising target absorbance values, and the Crystal Violet whole-cell staining method may be used to determine cell density so that absorbance values can be adjusted for plating differences.

MCM4 is a mini-chromosome maintenance protein that is essential for the initiation of eukaryotic genome replication and forms part of the pre-replication complex; the complex containing MCM4, MCM2, MCM6 and MCM7 possesses DNA helicase activity, and phosphorylation by CDC2 reduces this DNA helicase activity and chromatin binding. Dysregulation of MCM4 has been linked to diseases including cancer and developmental disorders, supporting its use as a biomarker for cell cycle research.