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      Alexa Fluor™ 647 Rat Anti-Mouse CD274 (PD-L1)
      Alexa Fluor™ 647 Rat Anti-Mouse CD274 (PD-L1)
      Multicolor flow cytometric analysis of CD274 (PD-L1) expression on resting and activated Mouse splenocytes. BALB/c mouse splenic leucocytes were either not activated (Upper Plots) or activated (Lower Plots) by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 553057) for 3 days (37°C). The cells were harvested, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse CD4 antibody (Cat. No. 553049) and with either Alexa Fluor™ 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; Left Plots) or Alexa Fluor™ 647 Rat Anti-Mouse CD274 (PD-L1) antibody (Cat. No. 568305/568306; Right Plots) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plots showing the correlated expression of CD274 (PD-L1) [or Ig Isotype control staining] versus CD4 were derived from gated events with the forward and side light- scatter characteristics of viable (DAPI-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
      Alexa Fluor™ 647 Rat Anti-Mouse CD274 (PD-L1)
      Multicolor flow cytometric analysis of CD274 (PD-L1) expression on resting and activated Mouse splenocytes. BALB/c mouse splenic leucocytes were either not activated (Upper Plots) or activated (Lower Plots) by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 553057) for 3 days (37°C). The cells were harvested, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse CD4 antibody (Cat. No. 553049) and with either Alexa Fluor™ 647 Rat IgG2b, κ Isotype Control (Cat. No. 557691; Left Plots) or Alexa Fluor™ 647 Rat Anti-Mouse CD274 (PD-L1) antibody (Cat. No. 568305/568306; Right Plots) at 0.5 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The bivariate pseudocolor density plots showing the correlated expression of CD274 (PD-L1) [or Ig Isotype control staining] versus CD4 were derived from gated events with the forward and side light- scatter characteristics of viable (DAPI-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 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™
      Cd274; Pdl1; Pdcd1l1; Pdcd1lg1; B7 homolog 1; B7h1
      Mouse (QC Testing)
      Rat LEW, also known as Lewis IgG2b, κ
      Mouse PD-L1
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      Aqueous buffered solution containing ≤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 the dye under optimum conditions and unreacted dye was removed.
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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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      4. An isotype control should be used at the same concentration as the antibody of interest.
      5. 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.
      6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      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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      568306 Rev. 2
      Antibody Details
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      10F.9G2

      The 10F.9G2 monoclonal antibody specifically recognizes Programmed cell death 1 ligand 1 (PDCD1 ligand 1) which is also known as Programmed death ligand 1 (PD-L1), B7 homolog 1 (B7-H1), or CD274. CD274 (PD-L1) is a 43-kDa type I transmembrane glycoprotein that is encoded by Cd274 which belongs to the B7 family within the Ig superfamily. CD274 (PD-L1) is expressed at low levels on resting peripheral T and B lymphocytes, monocytes, macrophages, dendritic cells (DC), and NK cells and undergoes upregulated expression upon cellular activation. PD-L1 (CD274) is also widely expressed on nonhematopoietic cell types including epithelial cells, endothelial cells, placental trophoblasts, and tumor cells. CD274 (PD-L1) and CD273 (PD-L2) serve as ligands for the CD279 (Programmed cell death protein 1/PD-1) inhibitory receptor. CD274 (PD-L1)-mediated signaling through CD279 (PD-1) regulates T cell responses in lymphoid and nonlymphoid tissues that are important for ensuring protective immunity while maintaining peripheral tolerance. This immune signaling checkpoint may suppress antitumor immune responses and prevent tumor rejection. The 10F.9G2 antibody reportedly blocks CD274 (PD-L1) binding to CD279 (PD-1) and can enhance the proliferation and effector responses of activated T cells including cytokine production and cytolytic activity.

      568306 Rev. 2
      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
      568306 Rev.2
      Citations & References
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      View product citations for antibody "568306" on CiteAb

      Development References (6)

      1. Eppihimer MJ, Gunn J, Freeman GJ, et al. Expression and regulation of the PD-L1 immunoinhibitory molecule on microvascular endothelial cells. Microcirculation. 2002; 9(2):133-145. (Immunogen: ELISA, Flow cytometry, Immunohistochemistry, Radioimmunoassay). View Reference
      2. Freeman GJ, Long AJ, Iwai Y, et al. Engagement of PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation. J Exp Med. 2000; 192:1027-1034. (Biology). View Reference
      3. Iraolagoitia XL, Spallanzani RG, Torres NI, et al. NK Cells Restrain Spontaneous Antitumor CD8+ T Cell Priming through PD-1/PD-L1 Interactions with Dendritic Cells.. J Immunol. 2016; 197(3):953-61. (Clone-specific: Flow cytometry). View Reference
      4. Paterson AM, Brown KE, Keir ME, et al. The programmed death-1 ligand 1:B7-1 pathway restrains diabetogenic effector T cells in vivo.. J Immunol. 2011; 187(3):1097-105. (Clone-specific: Blocking). View Reference
      5. Rodig N, Ryan T, Allen JA, et al. Endothelial expression of PD-L1 and PD-L2 down-regulates CD8+ T cell activation and cytolysis.. Eur J Immunol. 2003; 33(11):3117-26. (Clone-specific: Blocking, Flow cytometry, Functional assay). View Reference
      6. Sharpe AH, Wherry EJ, Ahmed R, Freeman GJ. The function of programmed cell death 1 and its ligands in regulating autoimmunity and infection.. Nat Immunol. 2007; 8(3):239-45. (Biology). View Reference
      View All (6) View Less
      568306 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.