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      PE Rat Anti-Mouse CD273
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      Consider BD Horizon RealYellow™ 586 (RY586) Reagents, a bright and clean fluorochrome alternative to PE off the yellow-green laser. RY586 can be used alongside PE on spectral cytometers. More Info #
      PE Rat Anti-Mouse CD273
      Splenic DC express CD273. Splenic DC were generated from BALB/c mice by culture with recombinant mouse GM-CSF (Cat. No. 554586). The cells were stained with either PE Rat anti-mouse B7-DC mAb TY25 (open histograms) or PE Rat IgG2a, κ Isotype control mAb R35-95 (Cat. No. 554689, filled histograms) in the presence of Mouse BD Fc Block™ Purified Anti-Mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142).  DC were identified by staining with FITC Rat Anti-Mouse CD11b mAb M1/70 (Cat. No. 553310/557396) and APC Hamster Anti-Mouse CD11c mAb HL3 (Cat. No. 550261).  The left panel displays CD273 expression on CD11b-positive CD11c-positive DC, while the right panel displays CD273 on CD11b-positive CD11c-negative DC.  CD11b-negative CD11c-negative non-DC had almost no CD273 expression (data not shown).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
      PE Rat Anti-Mouse CD273
      Splenic DC express CD273. Splenic DC were generated from BALB/c mice by culture with recombinant mouse GM-CSF (Cat. No. 554586). The cells were stained with either PE Rat anti-mouse B7-DC mAb TY25 (open histograms) or PE Rat IgG2a, κ Isotype control mAb R35-95 (Cat. No. 554689, filled histograms) in the presence of Mouse BD Fc Block™ Purified Anti-Mouse CD16/CD32 mAb 2.4G2 (Cat. No. 553141/553142).  DC were identified by staining with FITC Rat Anti-Mouse CD11b mAb M1/70 (Cat. No. 553310/557396) and APC Hamster Anti-Mouse CD11c mAb HL3 (Cat. No. 550261).  The left panel displays CD273 expression on CD11b-positive CD11c-positive DC, while the right panel displays CD273 on CD11b-positive CD11c-negative DC.  CD11b-negative CD11c-negative non-DC had almost no CD273 expression (data not shown).  Flow cytometry was performed on a BD FACSCalibur™ flow cytometry system.
      Product Details
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      BD Pharmingen™
      Pdcd1lg2; B7dc; B7-DC; PD-1 ligand 2; PD-L2; Btdc
      Mouse (QC Testing)
      Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
      Mouse B7-DC
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      58205
      AB_396874
      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 unconjugated antibody and free dye were 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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      3. An isotype control should be used at the same concentration as the antibody of interest.
      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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      6. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      7. For U.S. patents that may apply, see bd.com/patents.
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      557796 Rev. 2
      Antibody Details
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      TY25

      The TY25 monoclonal antibody specifically binds to CD273, also known as B7-DC or PD-L2. CD273 is a 42-kDa type I membrane glycoprotein encoded by the Pdcd1lg2 gene of the B7 family of the Ig superfamily. Pdcd1lg2 mRNA is expressed selectively in dendritic cells (DC), liver, and a few transformed cell lines (hepatic, neuroblastoma, and myeloid), but not in lymphoid tissues or macrophages. The protein has not been detected on resting peripheral T and B lymphocytes, macrophages, or DC, but it is upregulated upon activation of T cells, macrophages, and DC by a variety of stimulatory factors. B7-DC's receptor, PD-1, contains an ITIM (Immunoreceptor Tyrosine-based Inhibitory Motif) on its intracytoplasmic region and is expressed on activated B and T lymphocytes, suggesting that B7-DC-PD-1 interaction may be involved in the negative regulation of immune responses. The second PD-1 ligand, B7-H1 (PD-L1), is also a member of the B7 family of the Ig superfamily.  B7-DC may also participate in positive immunoregulation, or costimulation of T cells, through an additional receptor, which is not PD-1 and distinct from the alternate receptor for B7-H1. Moreover, B7-DC is the first B7 ligand that has been found to participate in bi-directional communication; ligation of B7-DC on DC stimulates those DC. The TY25 antibody blocks the binding of B7-DC to PD-1, but not to its alternate ligand.

      557796 Rev. 2
      Format Details
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      PE
      R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
      altImg
      PE
      Yellow-Green 488 nm, 532 nm, 561 nm
      496 nm, 566 nm
      576 nm
      557796 Rev.2
      Citations & References
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      Development References (11)

      1. Ansari MJ, Salama AD, Chitnis T, et al. The programmed death-1 (PD-1) pathway regulates autoimmune diabetes in nonobese diabetic (NOD) mice. J Exp Med. 2003 July; 198(1):63-69. (Clone-specific). View Reference
      2. Carreno BM, Collins M. The B7 family of ligands and its receptors: New pathways for costimulation and inhibition of immune responses. Annu Rev Immunol. 2002; 20:29-53. (Biology). View Reference
      3. Latchman Y, Wood CR, Chernova T, et al. PD-L2 is a second ligand for PD-1 and inhibits T cell activation. Nat Immunol. 2001; 2(3):261-268. (Biology). View Reference
      4. Liu X, Gao JX, Wen J, et al. B7DC/PDL2 promotes tumor immunity by a PD-1-independent mechanism. J Exp Med. 2003; 197:1721-1730. (Clone-specific). View Reference
      5. Nguyen LT, Radhakrishnan S, Ciric B, et al. Cross-linking the B7 family molecule B7-DC directly activates immune functions of dendritic cells. J Exp Med. 2002; 196:1393-1398. (Biology). View Reference
      6. Salama AD, Chitnis T, Imitola J, et al. Critical role of the programmed death-1 (PD-1) pathway in regulation of experimental autoimmune encephalomyelitis. J Exp Med. 2003 July; 198(1):71-78. (Biology). View Reference
      7. Shin T, Kennedy G, Gorski K et al. Cooperative B7-1/2 (CD80/CD86) and B7-DC costimulation of CD4+ T cells independent of the PD-1 receptor. J Exp Med. 2003; 198:31-38. (Biology). View Reference
      8. Tseng S-Y, Otsuji M, Gorski K, et al. B7-DC, a new dendritic cell molecule with potent costimulatory properties for T cells. J Exp Med. 2001; 193:839-846. (Biology). View Reference
      9. Tsushima F, Iwai H, Otsuki N, et al. Preferential contribution of B7-H1 to programmed death-1-mediated regulation of hapten-specific allergic inflammatory responses. Eur J Immunol. 2003; 33(10):2773-2782. (Clone-specific). View Reference
      10. Wang S, Bajorath J, Flies DB, Dong H, Honjo T, Chen L. Molecular modeling and functional mapping of B7-H1 and B7-DC uncouple costimulatory function from PD-1 interaction. J Exp Med. 2003; 197:1083-1091. (Biology). View Reference
      11. Yamazaki T, Akiba H, Iwai H, et al. Expression of programmed death 1 ligands by murine T cells and APC. J Immunol. 2002; 169(10):5538-5545. (Immunogen). View Reference
      View All (11) View Less
      557796 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.

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