eNOS Colorimetric Cell-Based ELISA

This in-cell ELISA kit from Assay Genie provides a high-throughput, lysate-free method for measuring eNOS (Endothelial Nitric Oxide Synthase) protein levels in cell lysates and cell culture supernatants. The kit can also be used to screen for the effects of various treatments, inhibitors (such as siRNA or chemicals), or activators on eNOS. eNOS is a key enzyme in the production of nitric oxide, a signaling molecule involved in regulating blood vessel function and cardiovascular health, and its dysregulation has been implicated in various cardiovascular diseases.

Reactivity has been demonstrated with human, mouse, and rat samples, and the kit uses a colorimetric detection method read at 450 nm. The assay has a dynamic range of > 5000 Cells and is supplied in a 96-well microplate format. The kit is intended for use with adherent cells and is recommended for storage at 4°C with 6 months of stability.

Detection is achieved through an indirect ELISA format in which eNOS-specific primary antibodies are bound by HRP-conjugated secondary antibodies; HRP activity is then revealed by a colorimetric substrate reaction. Because the assay is qualitative, two normalization approaches are provided: a monoclonal anti-GAPDH antibody serves as an internal positive control, and Crystal Violet whole-cell staining can be used to determine cell density and adjust for plating differences.