Purification notesWhole antiserum was fractionated and then further purified by ion-exchange chromatography to provide the IgG fraction of antiserum which is essentially free of other rabbit serum proteins.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/10000. Predicted molecular weight: 133 kDa.
1/100. Fix with acetone.
Use at an assay dependent concentration. PubMed: 20702812
FunctionProduces nitric oxide (NO) which is implicated in vascular smooth muscle relaxation through a cGMP-mediated signal transduction pathway. NO mediates vascular endothelial growth factor (VEGF)-induced angiogenesis in coronary vessels and promotes blood clotting through the activation of platelets. Isoform eNOS13C: Lacks eNOS activity, dominant-negative form that may down-regulate eNOS activity by forming heterodimers with isoform 1.
Tissue specificityPlatelets, placenta, liver and kidney.
Involvement in diseaseVariation in NOS3 seem to be associated with susceptibility to coronary spasm.
Sequence similaritiesBelongs to the NOS family. Contains 1 FAD-binding FR-type domain. Contains 1 flavodoxin-like domain.
Post-translational modificationsPhosphorylation by AMPK at Ser-1177 in the presence of Ca(2+)-calmodulin (CaM) activates activity. In absence of Ca(2+)-calmodulin, AMPK also phosphorylates Thr-495, resulting in inhibition of activity (By similarity). Phosphorylation of Ser-114 by CDK5 reduces activity.
Cellular localizationCell membrane. Membrane, caveola. Cytoplasm, cytoskeleton. Golgi apparatus. Specifically associates with actin cytoskeleton in the G2 phase of the cell cycle and which is favored by interaction with NOSIP and results in a reduced enzymatic activity.