ZIC1/2/3 Colorimetric Cell-Based ELISA

From Assay Genie, this Cell-Based ELISA kit provides a high-throughput, lysate-free method for detecting the ZIC1, ZIC2, and ZIC3 proteins, members of the ZIC family of C2H2-type zinc finger transcription factors. The assay uses an indirect ELISA format, in which target-specific primary antibodies capture ZIC1/2/3 in cultured cells and HRP-conjugated secondary antibodies bind the Fc region of the primary antibody, enabling a colorimetric reaction upon substrate addition. The kit is supplied in a 96-Well Microplate format and is suited to research in developmental biology, where the ZIC family plays critical roles in neural tube formation, cell differentiation, and organ development.

The assay is read at 450 nm using a standard microplate reader and supports a dynamic range of > 5000 Cells. It is reactive with human and mouse samples, with adherent cells as the specified sample type. The kit is stored at 4 °C with a stability of 6 months. Researchers can use the kit to measure relative ZIC1/2/3 levels in cultured cells and to screen the effects of treatments, inhibitors (such as siRNA or chemicals), or activators on ZIC1/2/3 expression.

Because the Cell-Based ELISA is qualitative, two normalization approaches are provided. 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 can be used to determine cell density, allowing absorbance values to be normalized to cell amounts and plating differences to be adjusted.