SKP1A/p19 Colorimetric Cell-Based ELISA Kit

From Assay Genie, this In-Cell ELISA Kit is designed for the accurate detection of SKP1A and P19 levels in cell lysates and culture supernatants. The cell-based format uses a colorimetric detection method at 450 nm and is reactive with human, mouse, and rat samples, making it suitable for adherent cell analysis. With a dynamic range of > 5000 Cells, the assay offers high throughput and sensitive monitoring of SKP1/p19 protein expression profiles in cultured cells, allowing researchers to measure relative amounts of SKP1/p19 and to screen the effects of various treatments, inhibitors (such as siRNA or chemicals), and activators.

Qualitative determination of SKP1A/p19 concentration is achieved through an indirect ELISA format, in which SKP1A/p19-specific primary antibodies capture the target while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody. The HRP enzyme then catalyzes a colorimetric reaction upon substrate addition. Because the assay is qualitative, normalization methods are required: a monoclonal antibody specific for human GAPDH serves as an internal positive control for normalizing target absorbance values, and following colorimetric measurement, a Crystal Violet whole-cell staining method can be used to determine cell density and adjust for plating differences.

SKP1A is an essential component of the SCF (SKP1-CUL1-F-box protein) ubiquitin ligase complex, which mediates the ubiquitination of proteins involved in cell cycle progression, signal transduction, and transcription. The kit is intended for storage at 4°C with 6 months stability.