Cell Signaling Technology

Product Pathways - Akt Signaling

Phospho-eNOS (Thr495) Antibody #9574

Applications Reactivity Sensitivity MW (kDa) Source
W IP Pg B (H) (M) (R) Endogenous 140 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Pg=Pig  B=Bovine
Species cross-reactivity is determined by Western blot.

Specificity / Sensitivity

Phospho-eNOS (Thr495) Antibody detects endogenous levels of eNOS only when phosphorylated at threonine 495.

Source / Purification

Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr495 of human eNOS. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from porcine aortic endothelial cells (PAECs), 3 days post-confluence, untreated, H2O2-treated (0.3 mM for 30 minutes) or calf intestinal alkaline phosphatase (CIP)-treated as indicated, using Phospho-eNOS (Thr495) Antibody (upper) or control eNOS antibody (lower).

Background

Endothelial nitric-oxide synthase (eNOS) is an important enzyme in the cardiovascular system. It catalyzes the production of nitric oxide (NO), a key regulator of blood pressure, vascular remodeling and angiogenesis (1,2). The activity of eNOS is regulated by phosphorylation at multiple sites. The two most thoroughly studied sites are the activation site Ser1177 and the inhibitory site Thr495 (3). Several protein kinases including Akt/PKB, PKA and AMPK activate eNOS by phosphorylating Ser1177 in response to various stimuli (4,5). In contrast, bradykinin and hydrogen peroxide activate eNOS activity by promoting Thr495 dephosphorylation (6,7).

  1. Fulton, D. et al. (2001) J. Pharmacol. Exp. Ther. 299, 818-824.
  2. Shaul, P.W. (2002) Annu. Rev. Physiol. 64, 749-774.
  3. Chen, Z.P. et al. (1999) FEBS Lett. 443, 285-289.
  4. Dimmeler, S. et al. (1999) Nature 399, 601-605.
  5. Fulton, D. et al. (1999) Nature 399, 597-601.
  6. Harris, M.B. et al. (2001) J. Biol. Chem. 276, 16587-16591.
  7. Thomas, S.R. et al. (2002) J. Biol. Chem. 277, 6017-6024.

Application References

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Companion Products

This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.

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