NEDD8 Colorimetric Cell-Based ELISA Kit

From Assay Genie, this cell-based ELISA kit is designed for the qualitative detection of NEDD8 protein expression profiles in cultured cells, using an indirect colorimetric format. The assay is described as lysate-free, high throughput, and sensitive, suitable for measuring relative NEDD8 levels in cells and for screening the effects of treatments, inhibitors (including siRNA and chemicals), and activators. Reactivity covers human, mouse, and rat samples, with detection by colorimetry at 450 nm and a dynamic range of greater than 5000 cells.

The workflow uses NEDD8-specific primary antibodies to capture the target, with HRP-conjugated secondary antibodies binding the Fc region of the primary antibody to catalyse a colorimetric reaction upon substrate addition. A monoclonal anti-GAPDH antibody is included as an internal positive control for normalising target absorbance values, and a Crystal Violet whole-cell staining method may be used to determine cell density. The kit is positioned for signal transduction research, with applications in studies of cell growth, differentiation, and stress response, and is stored at 4°C for 6 months.

NEDD8 is a member of the ubiquitin family of proteins, approximately 60% identical to ubiquitin, activated by an E1-like complex of App-b1 and Uba3 and then linked to the E2-like enzyme Ubc12. The major target protein modified by nedd8 is cullin-4a, and the protein is reported to be down-regulated during the development of brain.