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      Alexa Fluor™ 647 Rat Anti-Mouse iNOS
      Alexa Fluor™ 647 Rat Anti-Mouse iNOS
      Multiparameter flow cytometric analysis of iNOS expression in stimulated C57BL/6 Mouse thioglycolate-elicited peritoneal exudate cells. C57BL/6 thioglycollate-elicited peritoneal cells (Thio-PECs) were stimulated with Recombinant Mouse IFN-γ protein (Cat. No. 554587) and lipopolysaccharide (LPS) overnight (37°C). Cells were harvested, washed, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), washed, and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash Buffer with BD Horizon™ BUV395 Rat Anti-Mouse F4/80 antibody (Cat. No. 565614) and with either Alexa Fluor™ 647 Rat IgG2b, κ Isotype Control (Cat. No. 571480; Left Plot) or Alexa Fluor™ 647 Rat Anti-Mouse iNOS antibody (Cat. No. 572305/572306; Right Plot) at 0.25 µg/test. The bivariate pseudocolor density plot showing the correlated expression of iNOS (or Ig Isotype control staining) versus F4/80 was derived from gated events with the forward and side-light scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      Alexa Fluor™ 647 Rat Anti-Mouse iNOS
      Multiparameter flow cytometric analysis of iNOS expression in stimulated C57BL/6 Mouse thioglycolate-elicited peritoneal exudate cells. C57BL/6 thioglycollate-elicited peritoneal cells (Thio-PECs) were stimulated with Recombinant Mouse IFN-γ protein (Cat. No. 554587) and lipopolysaccharide (LPS) overnight (37°C). Cells were harvested, washed, fixed with BD Cytofix™ Fixation Buffer (Cat. No. 554655), washed, and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained in BD Perm/Wash Buffer with BD Horizon™ BUV395 Rat Anti-Mouse F4/80 antibody (Cat. No. 565614) and with either Alexa Fluor™ 647 Rat IgG2b, κ Isotype Control (Cat. No. 571480; Left Plot) or Alexa Fluor™ 647 Rat Anti-Mouse iNOS antibody (Cat. No. 572305/572306; Right Plot) at 0.25 µg/test. The bivariate pseudocolor density plot showing the correlated expression of iNOS (or Ig Isotype control staining) versus F4/80 was derived from gated events with the forward and side-light scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ Cell Analyzer System and FlowJo™ Software. Data shown on this Technical Data Sheet are not lot specific.
      Product Details
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      BD Pharmingen™
      iNOS; i-NOS; Nos2; Nos-2; MAC-NOS
      Mouse (QC Testing)
      Rat IgG2b, κ
      Mouse iNOS
      Intracellular staining (flow cytometry) (Routinely Tested)
      0.2 mg/ml
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated to the dye under optimum conditions and unreacted dye was removed. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
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      Recommended Assay Procedures

      BD® CompBeads can be used as surrogates to assess fluorescence spillover (compensation).  When fluorochrome conjugated antibodies are bound to BD® CompBeads, they have spectral properties very similar to cells.  However, for some fluorochromes there can be small differences in spectral emissions compared to cells, resulting in spillover values that differ when compared to biological controls.  It is strongly recommended that when using a reagent for the first time, users compare the spillover on cell and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
      2. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
      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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. An isotype control should be used at the same concentration as the antibody of interest.
      7. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
      8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      9. Alexa Fluor™ is a trademark of Life Technologies Corporation.
      10. For U.S. patents that may apply, see bd.com/patents.
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      572305 Rev. 1
      Antibody Details
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      CC02-30

      The CC02-30 monoclonal antibody specifically recognizes inducible nitric oxide synthase (iNOS) which is also known as NOS type II  (Nos-2 or NOS-II) or Macrophage NOS (MAC-NOS). This enzyme is encoded by Nos2 (nitric oxide synthase 2, inducible). The iNOS homodimer catalyzes the synthesis of nitric oxide (NO) in macrophages and other cell types that are induced to express iNOS in response to certain cytokines (eg, IFN-γ, TNF-α, and IL-1) and microbial products (eg, lipopolysaccharides, proteins, nucleic acids). iNOS acts through a multistep process to catalyze the generation of nitric oxide from L-arginine using molecular oxygen and NADPH as an electron donor. The NO free radical is highly reactive with other molecules such as proteins and nucleic acids of microbial origin and plays an important role in immunity by killing microbial pathogens to help counter infections. iNOS belongs to the NOS family that includes two other different nitric oxide synthases (isozymes): i) Neuronal NOS (nNOS encoded by Nos1) expressed in neurons and involved in neurotransmission as well as ii) Endothelial NOS (eNOS encoded by Nos3) expressed by endothelial cells and involved in regulating blood flow and vascular tone.

      572305 Rev. 1
      Format Details
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      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 670-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
      572305 Rev.1
      Citations & References
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      View product citations for antibody "572305" on CiteAb

      Development References (7)

      1. Bogdan C. Nitric oxide and the immune response.. Nat Immunol. 2001; 2(10):907-16. (Biology). View Reference
      2. Chen LC, Pace JL, Russell SW, Morrison DC. Altered regulation of inducible nitric oxide synthase expression in macrophages from senescent mice. Infect Immun. 1996; 64(10):4288-4298. (Biology). View Reference
      3. Landes MB, Rajaram MV, Nguyen H, Schlesinger LS. Role for NOD2 in Mycobacterium tuberculosis-induced iNOS expression and NO production in human macrophages.. J Leukoc Biol. 2015; 97(6):1111-9. (Biology). View Reference
      4. Lee SH, Kwak CH, Lee SK, et al. Anti-Inflammatory Effect of Ascochlorin in LPS-Stimulated RAW 264.7 Macrophage Cells Is Accompanied With the Down-Regulation of iNOS, COX-2 and Proinflammatory Cytokines Through NF-κB, ERK1/2, and p38 Signaling Pathway.. J Cell Biochem. 2016; 117(4):978-87. (Biology). View Reference
      5. Salim T, Sershen CL, May EE. Investigating the Role of TNF-α and IFN-γ Activation on the Dynamics of iNOS Gene Expression in LPS Stimulated Macrophages.. PLoS One. 2016; 11(6):e0153289. (Biology). View Reference
      6. Suschek CV, Schnorr O, Kolb-Bachofen V. The role of iNOS in chronic inflammatory processes in vivo: is it damage-promoting, protective, or active at all?. Curr Mol Med. 2004; 4(7):763-75. (Biology). View Reference
      7. Takada Y, Ye X, Simon S. The integrins.. Genome Biol. 2007; 8(5):215. (Biology). View Reference
      View All (7) View Less
      572305 Rev. 1

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

       

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      For Research Use Only. Not for use in diagnostic or therapeutic procedures.