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BD Transduction Laboratories™ Purified Mouse Anti-eNOS (pT495)
Clone 31/eNOS(pT495) (RUO)
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
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
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
612707 Rev. 1
Antibody Details
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
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
View product citations for antibody "612707" on CiteAb
Development References (1)
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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
612707 Rev. 1
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Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.