eNOS (Phospho-Thr494)Colorimetric Cell-Based ELISA Kit

Assay Genie offers this Cell-Based ELISA designed for the precise detection of phosphorylated eNOS (Endothelial Nitric Oxide Synthase) levels in cell lysates, with the kit specifically targeting the Phospho-Thr494 site. The assay is validated for use with Human, Mouse, and Rat samples using adherent cells. Detection is achieved through a Colorimetric method at 450 nm, with a dynamic range of > 5000 Cells.

The kit employs a lysate-free, high-throughput format capable of detecting eNOS protein phosphorylation and expression profiles in cultured cells, making it suitable for screening the effects of various treatments, inhibitors, or activators on eNOS phosphorylation. It uses an indirect ELISA approach in which eNOS (Phospho-Thr494) is captured by specific primary antibodies, while HRP-conjugated secondary antibodies bind the Fc region of the primary antibody to catalyze a colorimetric reaction upon substrate addition. 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 method may be used to determine cell density for further normalization.

Phosphorylation of eNOS at Thr494 plays a significant role in regulating its activity, making this kit a relevant tool for studying endothelial function and cardiovascular diseases. eNOS is described as a constitutive nitric oxide synthase that synthesizes nitric oxide (NO) from arginine and oxygen, implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway. The associated gene symbol is NOS3, and the sub-type is Phospho.