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BV480 Mouse Anti-Human PD-L1 (CD274)
BV480 Mouse Anti-Human PD-L1 (CD274)
Flow cytometric analysis of PD-L1 (CD274) expression on activated Human lymphocytes. Human peripheral blood mononuclear cells were stimulated with Phytohemagglutinin (PHA) for 3 days and stained with either BD Horizon™ BV480 Mouse IgG2b, κ Isotype Control (Cat. No. 566080; dotted line histogram) or BD Horizon™ BV480 Mouse Anti-Human PD-L1 (CD274) antibody (Cat. No. 568622/568623; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) 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 (7-AAD-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 were stimulated with Phytohemagglutinin (PHA) for 3 days and stained with either BD Horizon™ BV480 Mouse IgG2b, κ Isotype Control (Cat. No. 566080; dotted line histogram) or BD Horizon™ BV480 Mouse Anti-Human PD-L1 (CD274) antibody (Cat. No. 568622/568623; solid line histogram). BD Via-Probe™ Cell Viability 7-AAD Solution (Cat. No. 555815/555816) 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 (7-AAD-negative) lymphocytes. 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™
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
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 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. 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. BD Horizon Brilliant Violet 480 is covered by one or more of the following US patents: 8,575,303; 8,354,239.
  6. 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.
  7. 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.
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
568622 Rev. 1
Antibody Details
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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.

568622 Rev. 1
Format Details
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BV480
The BD Horizon Brilliant Violet™ 480 (BV480) Dye is part of the BD Horizon Brilliant Violet™ family of dyes. This polymer-technology fluorochrome has an excitation maximum (Ex Max) of 440-nm and an emission maximum (Em Max) of 479-nm. Driven by BD innovation, BV480 is designed to be excited by the violet laser (405-nm) and detected using an optical filter centered near 480-nm (e.g., a 525/50 bandpass filter). The increased fluorescence intensity of BV480 and narrower emission spectra, make it a good alternative for BV510 or V500. Due to its excitation profile, BV480 will also has less cross-laser excitation with the UV laser, resulting in less spillover into UV channels compared to BV510. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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BV480
Violet 405 nm
440 nm
479 nm
568622 Rev.1
Citations & References
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View product citations for antibody "568622" on CiteAb

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
View All (5) View Less
568622 Rev. 1

 

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