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    Purified Mouse Anti-Human eNOS (pS633)

    BD Transduction Laboratories™ Purified Mouse Anti-Human eNOS (pS633)

    Clone 37/eNOS(S633)

    (RUO)
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    Purified Mouse Anti-Human eNOS (pS633)
    Western blot analysis for eNOS (pS633).  Human endothelial cells were either left untreated (lane 1) or were treated with 200 U/mL of lambda phosphatase for 1 hour at 37°C (lane 2). The top panel was probed with a mouse anti-eNOS antibody (Cat. No. 610296) and the bottom panel was probed with the mouse anti-eNOS (pS633) antibody at a 1:500 dilution with a band observable at ~ 140 kDa.
    Purified Mouse Anti-Human eNOS (pS633)
    Western blot analysis for eNOS (pS633).  Human endothelial cells were either left untreated (lane 1) or were treated with 200 U/mL of lambda phosphatase for 1 hour at 37°C (lane 2). The top panel was probed with a mouse anti-eNOS antibody (Cat. No. 610296) and the bottom panel was probed with the mouse anti-eNOS (pS633) antibody at a 1:500 dilution with a band observable at ~ 140 kDa.
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    BD Transduction Laboratories™
    Nitric Oxide Synthase
    Human (QC Testing)
    Mouse IgG1
    Phosphorylated Human eNOS (pS633) Peptide
    Western blot (Routinely Tested)
    140 kDa
    250 µg/ml
    AB_399909
    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.
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    Recommended Assay Procedures

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

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    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.
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    612665 Rev. 1
    Antibody Details
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    37/eNOS(S633)

    Nitirc 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 cytoxicity. 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-633 amd 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.

    612665 Rev. 1
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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
    612665 Rev.1
    Citations & References
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    Development References (4)

    1. Butt E, Bernhardt M, Smolenski A, et al. Endothelial nitric-oxide synthase (type III) is activated and becomes calcium independent upon phosphorylation by cyclic nucleotide-dependent protein kinases. J Biol Chem. 2000; 275(7):5179-5187. (Biology). View Reference
    2. Dimmeler S, Fleming I, Fisslthaler B, Hermann C, Busse R, Zeiher AM. Activation of nitric oxide synthase in endothelial cells by Akt-dependent phosphorylation.. Nature. 1999; 399(6736):601-605. (Biology). View Reference
    3. Gallis B, Corthals GL, Goodlett DR, et al. Identification of flow-dependent endothelial nitric-oxide synthase phosphorylation sites by mass spectrometry and regulation of phosphorylation and nitric oxide production by the phosphatidylinositol 3-kinase inhibitor LY294002. J Biol Chem. 1999; 274(42):30101-30108. (Biology). View Reference
    4. 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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    612665 Rev. 1

    Please refer to Support Documents for Quality Certificates


    Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


    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.

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