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RY586 Mouse Anti-Human PD-L2 (CD273)
RY586 Mouse Anti-Human PD-L2 (CD273)

Flow cytometric analysis of PD-L2 (CD273) expression on viable Human monocyte-derived mature dendritic cells.  Human monocyte-derived mature dendritic cells were stained with either BD Horizon™ RY586 Mouse IgG2a, κ Isotype Control (Cat. No. 568131; dashed line histogram) or BD Horizon™ RY586 Mouse Anti-Human PD-L2 (CD273) antibody (Cat. No. 568491/568492; solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing PD-L2 (CD273) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Flow cytometric analysis of PD-L2 (CD273) expression on viable Human monocyte-derived mature dendritic cells.  Human monocyte-derived mature dendritic cells were stained with either BD Horizon™ RY586 Mouse IgG2a, κ Isotype Control (Cat. No. 568131; dashed line histogram) or BD Horizon™ RY586 Mouse Anti-Human PD-L2 (CD273) antibody (Cat. No. 568491/568492; solid line histogram). DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. The fluorescence histogram showing PD-L2 (CD273) expression (or Ig Isotype control staining) was derived from gated events with the forward and side light-scatter characteristics of viable (DAPI-negative) cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Product Details
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BD Horizon™
CD273; PD-L2; PDL2; PDCD1LG2; B7-DC; B7DC
Human (QC Testing)
Mouse IgG2a, κ
Human PD-L2 cDNA
Flow cytometry (Routinely Tested)
5 µl
80380
AB_3684318
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.

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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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).
  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. Human donor specific background has been observed in relation to the presence of anti-polyethylene glycol (PEG) antibodies, developed as a result of certain vaccines containing PEG, including some COVID-19 vaccines. We recommend use of BD Horizon Brilliant™ Stain Buffer in your experiments to help mitigate potential background. For more information visit https://www.bdbiosciences.com/en-us/support/product-notices.
  7. CF™ is a trademark of Biotium, Inc.
  8. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  9. For U.S. patents that may apply, see bd.com/patents.
568491 Rev. 1
Antibody Details
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24F.10C12

The 24F.10C12 monoclonal antibody specifically recognizes Programmed Death Ligand 2 (PD-L2, PDL2) which is also known as CD273, and B7-DC (B7DC). PD-L2 (CD273) is a type 1 transmembrane glycoprotein that is encoded by PDCD1LG2 (programmed cell death 1 ligand 2) which belongs to the B7 family within the  Ig superfamily. PD-L2 (CD273) serves as a ligand for CD279, the Programmed Death 1 (PD-1) receptor. CD273 is expressed on dendritic cells, activated monocytes, medullary thymic epithelial cells, placental trophoblasts, and myocardial endothelium. PD-L2 (CD273) can function as a coinhibitor of T cell functions by binding to and signaling through CD279 (PD-1). PD-L2 (CD273) can also reportedly act through another receptor to costimulate T cell activation and cytokine production. The 24F.10C12 antibody can reportedly block the binding of PD-L2 (CD273) to CD279 (PD-1).

568491 Rev. 1
Format Details
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RY586
The BD Horizon RealYellow™ 586 (RY586) Dye is part of the BD family of yellow-green dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 565-nm and an emission maximum (Em Max) at 586-nm. Driven by BD innovation, RY586 can be used on both spectral and conventional cytometers and is designed to be excited by the Yellow-Green laser (561-nm) with minimal excitation by the 488-nm Blue laser. For conventional instruments equipped with a Yellow-Green laser (561-nm), RY586 can be used as an alternative to PE and we recommend using an optical filter centered near 586-nm (eg, a 586/15-nm bandpass filter). For spectral instruments equipped with a Yellow-Green laser (561-nm), it can be used in conjunction with PE. Compared to PE, RY586 is similar in brightness, minimal spillover into Blue detectors, and increased spillover into the 610/20-nm (PE-CF594) detector. Please ensure that your instrument configuration (lasers and optical filters) is appropriate for this dye.
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RY586
Yellow-Green 561 nm
564 nm
586 nm
568491 Rev.1
Citations & References
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View product citations for antibody "568491" on CiteAb

Development References (4)

  1. Brown JA, Dorfman DM, Ma FR, et al. Blockade of programmed death-1 ligand on dendritic cells enhances T cell activation and cytokine production. J Immunol. 2003; 170:1257-1266. (Immunogen: Blocking, Flow cytometry, Functional assay, Immunohistochemistry). View Reference
  2. Messal N, Serriari NE, Pastor S, Nunès JA, Olive D. PD-L2 is expressed on activated human T cells and regulates their function.. Mol Immunol. 2011; 48(15-16):2214-9. (Clone-specific: Flow cytometry, Stimulation). View Reference
  3. 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
  4. Tavukcuoglu E, Horzum U, Yilmaz KB, Esendagli G. PD-L2+ wound zone macrophage-like cells display M1/M2-mixed activation and restrain the effector Th1 responses.. Immunol Cell Biol. 2020; 98(2):152-164. (Clone-specific: Flow cytometry). View Reference
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568491 Rev. 1

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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.