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Purified Mouse Anti-eNOS (pT495)
Purified Mouse Anti-eNOS (pT495)
Western blot analysis of eNOS (pT495). A human endothelial cell lysate (lane 1),  recombinant eNOS protein before in vitro phosphorylation of Thr-495 (lane 2), and recombinant eNOS protein after in vitro phosphorylation of Thr-495 with PKA kinase (lane 3) was probed with a 1:1000 dilution of the mouse anti-eNOS (pT495) antibody. eNOS (pT495) may be observed to be migrating at ~ 140 kDa.
Western blot analysis of eNOS (pT495). A human endothelial cell lysate (lane 1),  recombinant eNOS protein before in vitro phosphorylation of Thr-495 (lane 2), and recombinant eNOS protein after in vitro phosphorylation of Thr-495 with PKA kinase (lane 3) was probed with a 1:1000 dilution of the mouse anti-eNOS (pT495) antibody. eNOS (pT495) may be observed to be migrating at ~ 140 kDa.
Product Details
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BD Transduction Laboratories™
Human (QC Testing), Mouse,Rat,Dog (Tested in Development)
Mouse IgG1
Human Phosphorylated eNOS Peptide
Western blot (Routinely Tested)
140 kDa
250 µg/ml
AB_399946
Aqueous buffered solution containing BSA, glycerol, and ≤0.09% sodium azide.
RUO


Preparation And Storage

The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at -20°C.

Recommended Assay Procedures

Western blot:  Please refer to http://www.bdbiosciences.com/pharmingen/protocols/Western_Blotting.shtml

Product Notices

  1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
  4. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
612707 Rev. 1
Antibody Details
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31/eNOS(pT495)

Nitric oxide synthase (NOS), a cell-type specific enzyme, catalyzes the synthesis of nitric oxide (NO). NO is a short-lived radical that transmits signals involved in vasorelaxation, neurotransmission, and cytotoxicity. In neurons and endothelial cells, constitutive NOS (cNOS) is activated by agonists that increase intracellular Ca2+ levels and enhance calmodulin binding. Neuronal NOS (nNOS) and  endothelial NOS (eNOS) have recognition sites for NADPH, FAD, FMN, and calmodulin. eNOS has a unique N-myristylation consensus sequence that may explain its membrane localization. Various protein kinases have been implicated in regulation of eNOS activity, including AMPK, PKA, PKB/Akt, PKC, and CaM Kinase II. During VEGF stimulation, eNOS is transiently phosphorylated at Ser-1177 by PKB/akt and dephosphorylated at Thr-495. At later time points, VEGF stimulation leads to an increase in Thr-495 phosphorylation mediated by PKC and a decrease in Ser-1177 phosphorylation. In addition, Ser-495, Ser-633, and Ser-1177 are phosphorylated by PKA and PKG in vitro. Thus, eNOS activity may be regulated through complex phosphorylation events mediated by multiple kinases at various phosphorylation sites.

612707 Rev. 1
Format Details
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Purified
Tissue culture supernatant is purified by either protein A/G or affinity purification methods. Both methods yield antibody in solution that is free of most other soluble proteins, lipids, etc. This format provides pure antibody that is suitable for a number of downstream applications including: secondary labeling for flow cytometry or microscopy, ELISA, Western blot, etc.
Purified
612707 Rev.1
Citations & References
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Development References (1)

  1. Michell BJ, Chen Zp, Tiganis T, et al. Coordinated control of endothelial nitric-oxide synthase phosphorylation by protein kinase C and the cAMP-dependent protein kinase. J Biol Chem. 2001; 276(21):17625-17628. (Biology). View Reference
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For Research Use Only. Not for use in diagnostic or therapeutic procedures.