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RB780 Mouse Anti-Human PD-L1 (CD274)
RB780 Mouse Anti-Human PD-L1 (CD274)
Flow cytometric analysis of PD-L1 (CD274) expression on activated Human lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated with Phytohemagglutinin (PHA) for 3 days and stained with either BD Horizon™ RB780 Mouse IgG2b, κ Isotype Control (Cat. No. 568741; dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Human PD-L1 (CD274) antibody (Cat. No. 569073/569074; 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 the expression of PD-L1 (CD274) [or Ig Isotype control staining] was derived from gated events with the forward and side light-scatter characteristics of activated viable (DAPI-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of PD-L1 (CD274) expression on activated Human lymphocytes. Human peripheral blood mononuclear cells (PBMC) were stimulated with Phytohemagglutinin (PHA) for 3 days and stained with either BD Horizon™ RB780 Mouse IgG2b, κ Isotype Control (Cat. No. 568741; dashed line histogram) or BD Horizon™ RB780 Mouse Anti-Human PD-L1 (CD274) antibody (Cat. No. 569073/569074; 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 the expression of PD-L1 (CD274) [or Ig Isotype control staining] was derived from gated events with the forward and side light-scatter characteristics of activated viable (DAPI-negative) lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
製品詳細
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BD Horizon™
B7H1; PD-L1; PDCD1L1; PDCD1LG1; PDL1; hPD-L1
Human (QC Testing)
Mouse BALB/c IgG2b, κ
Human PD-L1
Flow cytometry (Routinely Tested)
5 µl/test
29126
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.

推奨アッセイ手順

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. 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. 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. Please observe the following precautions: Absorption of visible light can significantly alter the energy transfer occurring in any tandem fluorochrome conjugate; therefore, we recommend that special precautions be taken (such as wrapping vials, tubes, or racks in aluminum foil) to prevent exposure of conjugated reagents, including cells stained with those reagents, to room illumination.
  6. 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.
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  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. Cy is a trademark of Global Life Sciences Solutions Germany GmbH or an affiliate doing business as Cytiva.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  11. For U.S. patents that may apply, see bd.com/patents.
569074 Rev. 2
抗体の詳細
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29E.2A3

The 29E.2A3 monoclonal antibody specifically recognizes Programmed cell death 1 ligand 1 (PDCD1 ligand 1, PDCD1L1, PDCD1LG1) which is also known as Programmed death ligand 1 (PD-L1 or PDL1) as well as CD274, or B7 homolog 1 (B7-H1, B7H1). PD-L1 (CD274) and PD-L2 (CD273) are type I transmembrane glycoproteins that belong to the B7 family within the Ig gene superfamily and serve as ligands for CD279 (Program Death 1/PD-1). PD-L1 (CD274) is expressed on antigen-presenting cells including activated monocytes, macrophages, and dendritic cells (DCs) as well as activated T cells, B cells, NK cells, and keratinocytes. PD-L1 (CD274) is also variably expressed on placental trophoblasts, myocardial endothelium, cortical thymic epithelial cells, and tumor cells. PD-L1-mediated signaling through PD-1 regulates T cell responses important for providing protective immunity while maintaining peripheral tolerance. This immune signaling checkpoint may also suppress antitumor immune responses and prevent tumor rejection. The 29E.2A3 antibody reportedly blocks PD-L1 (CD274) binding to CD279 (PD-1) and can enhance the proliferation and cytokine production of activated T cells.

569074 Rev. 2
フォーマットの詳細
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RB780
The BD Horizon RealBlue™ 780 (RB780) 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 781-nm. Driven by BD innovation, RB780 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), RB780 can be used as an alternative to PE-Cy7 and we recommend using an optical filter centered near 780-nm (eg, a 780/60-nm bandpass filter). For spectral instruments equipped with a Blue laser (488-nm), it can be used in conjunction with PE-Cy7. RB780 is on average brighter than PE-Cy7 and has minimal spillover into Yellow-Green detectors.
altImg
RB780
Blue 488 nm
498 nm
781 nm
569074 Rev.2
引用&参考文献
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Development References (5)

  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. (Clone-specific: Blocking, Enhancement, Flow cytometry, Immunohistochemistry). View Reference
  2. Butte MJ, Peña-Cruz V, Kim MJ, Freeman GJ, Sharpe AH. Interaction of human PD-L1 and B7-1.. Mol Immunol. 2008; 45(13):3567-72. (Clone-specific: Blocking, Flow cytometry). View Reference
  3. Grenga I, Donahue RN, Lepone LM, Richards J, Schlom J. A fully human IgG1 anti-PD-L1 MAb in an in vitro assay enhances antigen-specific T-cell responses.. Clin Transl Immunology. 2016; 5(5):e83. (Clone-specific: Flow cytometry). View Reference
  4. Hughes MJ, McGettrick HM, Sapey E. Importance of validating antibody panels: Anti-PD-L1 clone binds AF700 fluorophore. J Immunol Methods. 2020; 483:112795. (Clone-specific: Flow cytometry, Immunofluorescence). View Reference
  5. 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. (Immunogen: ELISA, Flow cytometry). View Reference
すべて表示する (5) 表示項目を減らす
569074 Rev. 2

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.