Skip to main content Skip to navigation
Hamburger Menu
Search icon
Country Selector Country Selector
Japan (Japanese)
Profile Icon Profile Icon
Sign-in/Register UserName
Products

Solutions

Discover & Learn

Resources & Tools

Support

Search icon Search icon
Close Menu
      Alert Icon
      Alexa Fluor® 647 Mouse anti-eNOS
      Alexa Fluor® 647 Mouse anti-eNOS
      Analysis of eNOS in human endothelial cells. EA-hy 926 cells (Edgell, McDonald, Graham, 1983) were either transfected with eNOS RNAi (open histogram) or untreated (shaded histogram).  The cells were fixed (BD Cytofix™ Fixation buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-eNOS.  Down-regulation of eNOS expression is evident in the RNAi-transfected cells.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.
      Alexa Fluor® 647 Mouse anti-eNOS
      Analysis of eNOS in human endothelial cells. EA-hy 926 cells (Edgell, McDonald, Graham, 1983) were either transfected with eNOS RNAi (open histogram) or untreated (shaded histogram).  The cells were fixed (BD Cytofix™ Fixation buffer, Cat. No. 554655) for 10 minutes at 37°C, then permeabilized (BD Phosflow™ Perm Buffer III, Cat. No. 558050) on ice for 30 minutes, and then stained with Alexa Fluor® 647 Mouse anti-eNOS.  Down-regulation of eNOS expression is evident in the RNAi-transfected cells.  Flow cytometry was performed on a BD™ LSR II flow cytometry system.
      製品詳細
      Down Arrow Up Arrow


      BD Phosflow™
      NOS type III, NOS3, EC-NOS, NOS III
      Human (QC Testing), Mouse (Reported)
      Mouse IgG1, κ
      Human eNOS aa. 1025-1203
      Intracellular staining (flow cytometry) (Routinely Tested)
      20 µl
      AB_1645410
      Aqueous buffered solution containing BSA and ≤0.09% sodium azide.
      RUO


      Preparation and Storage

      Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was removed.
      詳細を表示 blue down arrow 表示項目を減らす blue up arrow

      Product Notices

      1. This reagent has been pre-diluted for use at the recommended Volume per Test. We typically use 1 × 10^6 cells in a 100-µl experimental sample (a test).
      2. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      3. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
      4. 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.
      5. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      7. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
      8. An isotype control should be used at the same concentration as the antibody of interest.
      9. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      詳細を表示 blue down arrow 表示項目を減らす blue up arrow
      560102 Rev. 2
      抗体の詳細
      Down Arrow Up Arrow
      33/eNOS

      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 and both are regulated in a similar manner. The human forms exhibit 52% amino acid identity. However, they are distinct gene products of about 155 kDa (nNOS) and 140 kDa (eNOS). The eNOS gene was cloned from human vascular endothelium as well as from bovine aortic endothelial cells (BAEC). eNOS protein has a unique N-myristylation consensus sequence that may explain its membrane localization.

      The 33/eNOS monoclonal antibody recognizes eNOS, regardless of phosphorylation status.

      The specificity of this antibody conjugate for flow cytometric analysis was validated by confirming that RNA-mediated interference (RNAi) of the specific protein reduced the staining of the cells (see figure).  Furthermore, the capacity of the RNAi to down-regulate the expression of the relevant protein was confirmed by western blot analysis.

      560102 Rev. 2
      フォーマットの詳細
      Down Arrow Up Arrow
      Alexa Fluor™ 647
      Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      Alexa Fluor™ 647
      Red 627-640 nm
      653 nm
      669 nm
      560102 Rev.2
      引用＆参考文献
      Down Arrow Up Arrow

      Development References (6)

      1. Chen PF, Tsai AL, Wu KK. Cysteine 184 of endothelial nitric oxide synthase is involved in heme coordination and catalytic activity. J Biol Chem. 1994; 269(40):25062-25066. (Clone-specific: Western blot). View Reference
      2. Dinerman JL, Dawson TM, Schell MJ, Snowman A, Snyder SH. Endothelial nitric oxide synthase localized to hippocampal pyramidal cells: implications for synaptic plasticity. Proc Natl Acad Sci U S A. 1994; 91(10):4214-4218. (Biology). View Reference
      3. Edgell C-JS, McDonald CC, Graham JB. Permanent cell line expressing human factor VIII-related antigen established by hybridization. Proc Natl Acad Sci U S A. 1983; 80:3734-3737. (Methodology: Controls).
      4. Nathan C, Xie QW. Regulation of biosynthesis of nitric oxide. J Biol Chem. 1994; 269(19):13725-13728. (Biology). View Reference
      5. Shen YH, Zhang L, Utama B et al. Human cytomegalovirus inhibits Akt-mediated eNOS activation through upregulating PTEN (phosphatase and tensin homolog deleted on chromosome 10). Cardiovasc Res. 2006; 69(2):502-511. (Biology). View Reference
      6. Varghese P, Harrison RW, Lofthouse RA, Georgakopoulos D, Berkowitz DE, Hare JM. β3-adrenoceptor deficiency blocks nitric oxide-dependent inhibition of myocardial contractility. J Clin Invest. 2000; 106(5):697-703. (Clone-specific: Western blot). View Reference
      すべて表示する (6) 表示項目を減らす
      560102 Rev. 2

      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.

      Website Feedback