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      RB705 Rat Anti-Mouse CD273 (PD-L2)
      RB705 Rat Anti-Mouse CD273 (PD-L2)
      Multicolor flow cytometric analysis of CD273 (PD-L2) expression on viable Mouse splenic dendritic cells.  BALB/c Mouse splenic leukocytes were cultured overnight with Recombinant Mouse GM-CSF protein (Cat. No. 554586; 5 ng/ml). The cells were harvested and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Hamster Anti-Mouse CD11c antibody (Cat. No. 550261) and with either BD Horizon™ RB705 Rat IgG2a, κ Isotype Control (Cat. No. 570262; Left Plot) or BD Horizon™ RB705 Rat Anti-Mouse CD273 (PD-L2) antibody (Cat. No. 570284/570285; Right Plot) 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 plot showing the correlated expression of CD273 (PD-L2) [or Ig Isotype control staining] versus CD11c was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenic leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.​
      RB705 Rat Anti-Mouse CD273 (PD-L2)
      Multicolor flow cytometric analysis of CD273 (PD-L2) expression on viable Mouse splenic dendritic cells.  BALB/c Mouse splenic leukocytes were cultured overnight with Recombinant Mouse GM-CSF protein (Cat. No. 554586; 5 ng/ml). The cells were harvested and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with APC Hamster Anti-Mouse CD11c antibody (Cat. No. 550261) and with either BD Horizon™ RB705 Rat IgG2a, κ Isotype Control (Cat. No. 570262; Left Plot) or BD Horizon™ RB705 Rat Anti-Mouse CD273 (PD-L2) antibody (Cat. No. 570284/570285; Right Plot) 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 plot showing the correlated expression of CD273 (PD-L2) [or Ig Isotype control staining] versus CD11c was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) splenic leukocytes. Flow cytometry and data analysis were performed using a BD FACSymphony™ A5 SE Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.​
      Product Details
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      BD Horizon™
      B7-DC; Btdc; PD-1 ligand 2; PD-L2; PDCD1 ligand 2; programmed cell death 1 ligand 2
      Mouse (QC Testing)
      Rat SD, also known as Sprague-Dawley (outbred) IgG2a, κ
      Mouse B7-DC-transfected Cell Line
      Flow cytometry (Routinely Tested)
      0.2 mg/ml
      58205
      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 cells and BD® CompBeads to ensure that BD® CompBeads are appropriate for your specific cellular application.

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      Product Notices

      1. When using high concentrations of antibody, background binding of this dye to erythroid fragments produced by ammonium chloride-based lysis, such as with BD Pharm Lyse™ Lysing Buffer (Cat. No. 555899), has been observed when the antibody conjugate was present during the lysis procedure. This may cause nonspecific staining of target cells, such as leukocytes, which have bound the resulting erythroid fragments. This background can be mitigated by any of the following: titrating the antibody conjugate to a lower concentration, fixing samples with formaldehyde, or removing erythrocytes before staining (eg, gradient centrifugation or pre-lysis with wash). This background has not been observed when cells were lysed with BD FACS™ Lysing Solution (Cat. No. 349202) after staining.
      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. 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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      8. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
      9. For U.S. patents that may apply, see bd.com/patents.
      10. Please observe the following precautions: We recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to protect exposure of conjugated reagents, including cells stained with those reagents, to any room illumination. Absorption of visible light can significantly affect the emission spectra and quantum yield of tandem fluorochrome conjugates.
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      570284 Rev. 1
      Antibody Details
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      MIH37

      The MIH37 monoclonal antibody specifically recognizes Programmed cell death 1 ligand 2 (PD-L2) which is also known as CD273 and Butyrophilin B7-DC (B7-DC). This regulatory molecule is a 42-kDa type I membrane glycoprotein encoded by Pdcd1lg2 which belongs to the B7 family of the Ig superfamily. CD273 (PD-L2)  is comprised of an extracellular region with an N-terminal IgV-like domain followed by an IgC2-type domain, a transmembrane sequence, and a cytoplasmic tail. Although not detected on resting leucocytes, its expression is upregulated upon activation of macrophages and dendritic cells (DC) by a variety of stimulatory factors including IL-4, IL-13, or GM-CSF. CD273 (PD-L2) serves as a ligand for the coinhibitory receptor, CD279 (PD-1), a receptor that likewise binds to the CD274 (PD-L1) ligand. CD273 (PD-L2) also binds to Repulsive guidance molecule b (RGMb) that is expressed by T cells, macrophages, neutrophils, and DC. RGMb may associate with other receptors that transduce costimulatory or coinhibitory signals. The MIH37 antibody reportedly blocks the binding of CD273 (PB-L2) to its receptors, CD279 (PD-1) or RGMb and can inhibit antigen-specific T cell responses and hapten-induced contact hypersensitivity reactions. The TY25 monoclonal antibody reportedly binds with lower affinity near or at the same mouse CD273 (PD-L2) epitope recognized by the MIH37 antibody.

      570284 Rev. 1
      Format Details
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      RB705
      The BD Horizon RealBlue™ 705 (RB705) Dye is part of the BD® family of blue dyes. It is a tandem fluorochrome with an excitation maximum (Ex Max) at 498-nm and an emission maximum (Em Max) at 707-nm as measured using an antibody-dye conjugate. Driven by BD® innovation, RB705 can be used on both spectral and conventional cytometers and is designed to be excited by the Blue laser (488-nm) with minimal excitation by the 561-nm Yellow-Green laser. For conventional instruments equipped with a Blue laser (488-nm), RB705 can be used as an alternative to PerCP-Cy5.5 or BB700 and we recommend using an optical filter centered near 710-nm (e.g., a 695/40 or 710/50-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PerCP-Cy5.5. RB705 is on average brighter than PerCP-Cy5.5 and BB700, and has minimal spillover into Yellow-Green detectors.
      altImg
      RB705
      Blue 488 nm
      498 nm
      707 nm
      570284 Rev.1
      Citations & References
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      View product citations for antibody "570284" on CiteAb

      Development References (6)

      1. Matsumoto K, Fukuyama S, Eguchi-Tsuda M, et al. B7-DC induced by IL-13 works as a feedback regulator in the effector phase of allergic asthma.. Biochem Biophys Res Commun. 2008; 365(1):170-5. (Clone-specific: Flow cytometry). View Reference
      2. Nie X, Chen W, Zhu Y, et al. B7-DC (PD-L2) costimulation of CD4+ T-helper 1 response via RGMb.. Cell Mol Immunol. 2018; 15(10):888-897. (Biology). View Reference
      3. Okudaira K, Hokari R, Tsuzuki Y, et al. Blockade of B7-H1 or B7-DC induces an anti-tumor effect in a mouse pancreatic cancer model.. Int J Oncol. 2009; 35(4):741-9. (Clone-specific: Flow cytometry). View Reference
      4. Ritprajak P, Hashiguchi M, Akiba H, Yagita H, Okumura K, Azuma M. Antibodies against B7-DC with differential binding properties exert opposite effects.. Hybridoma (Larchmt). 2012; 31(1):40-7. (Immunogen: Flow cytometry). View Reference
      5. Xiao Y, Yu S, Zhu B, et al. RGMb is a novel binding partner for PD-L2 and its engagement with PD-L2 promotes respiratory tolerance.. J Exp Med. 2014; 211(5):943-59. (Clone-specific: Flow cytometry). View Reference
      6. 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. (Biology: Flow cytometry). View Reference
      View All (6) View Less
      570284 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.