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BUV395 Mouse Anti-Human PD-L2 (CD273)
BUV395 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™ BUV395 Mouse IgG2a, κ Isotype Control (Cat. No. 563809; dashed line histogram) or BD Horizon™ BUV395 Mouse Anti-Human PD-L2 (CD273) antibody (Cat. No. 569053/569054; 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™ BUV395 Mouse IgG2a, κ Isotype Control (Cat. No. 563809; dashed line histogram) or BD Horizon™ BUV395 Mouse Anti-Human PD-L2 (CD273) antibody (Cat. No. 569053/569054; 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/test
80380
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 unconjugated antibody and free dye were removed.

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.

For optimal and reproducible results, BD Horizon Brilliant™ Stain Buffer should be used anytime BD Horizon Brilliant™ dyes are used in a multicolor flow cytometry panel.  Fluorescent dye interactions may cause staining artifacts which may affect data interpretation.  The BD Horizon Brilliant Stain Buffer was designed to minimize these interactions. When BD Horizon Brilliant Stain Buffer is used in in the multicolor panel, it should also be used in the corresponding compensation controls for all dyes to achieve the most accurate compensation. For the most accurate compensation, compensation controls created with either cells or beads should be exposed to BD Horizon Brilliant Stain Buffer for the same length of time as the corresponding multicolor panel. More information can be found in the Technical Data Sheet of the BD Horizon Brilliant Stain Buffer (Cat. No. 563794/566349) or the BD Horizon Brilliant Stain Buffer Plus (Cat. No. 566385).

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. 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).
  4. An isotype control should be used at the same concentration as the antibody of interest.
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. BD Horizon Brilliant Ultraviolet 395 is covered by one or more of the following US patents: 8,158,444; 8,575,303; 8,354,239.
  7. BD Horizon Brilliant Stain Buffer is covered by one or more of the following US patents: 8,110,673; 8,158,444; 8,575,303; 8,354,239.
  8. 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.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. For U.S. patents that may apply, see bd.com/patents.
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).

Format Details
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BUV395
The BD Horizon Brilliant™ Ultraviolet 395 (BUV395) Dye is part of the BD Horizon Brilliant™ Ultraviolet family of dyes. This base dye is a polymer fluorochrome with an excitation maximum (Ex Max) of 348-nm and an emission maximum (Em Max) at 395-nm. Driven by BD innovation, BUV395 is designed to be excited by the ultraviolet laser (355-nm) and detected using an optical filter centered near 380-nm (e.g., 379/28-nm bandpass filter). BUV395 is the ideal dye when using only one detector on the ultraviolet laser as it spills into no other detectors and no other fluors spill into it. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BUV395
Ultraviolet 355 nm
348 nm
395 nm
Citations & References
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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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