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BV480 Rat Anti-Mouse CD274 (PD-L1)
BV480 Rat Anti-Mouse CD274 (PD-L1)
Multicolor flow cytometric analysis of CD274 (PD-L1) expression on resting and activated Mouse splenocytes. BALB/c Mouse splenic leucocytes were either not activated (Top Plots) or activated (Bottom Plots) by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 553057/567115/567114) for 3 days (37°C). The cells were harvested, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse CD4 antibody (Cat. No. 553049/553048/561837) and with either BD Horizon™ BV480 Rat IgG2b, κ Isotype Control (Cat. No. 565649; Left Plots) or BD Horizon™ BV480 Rat Anti-Mouse CD274 (PD-L1) antibody (Cat. No. 568589/568590; Right Plots) at 1.0 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of CD274 (PD-L1) [or Ig Isotype control staining] versus CD4 were derived from gated events with the forward and side light scatter characteristics of viable (DAPI-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Multicolor flow cytometric analysis of CD274 (PD-L1) expression on resting and activated Mouse splenocytes. BALB/c Mouse splenic leucocytes were either not activated (Top Plots) or activated (Bottom Plots) by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3e antibody (Cat. No. 553057/567115/567114) for 3 days (37°C). The cells were harvested, washed and preincubated with Purified Rat Anti-Mouse CD16/CD32 antibody (Mouse BD Fc Block™) [Cat. No. 553141/553142]. The cells were then stained with PE Rat Anti-Mouse CD4 antibody (Cat. No. 553049/553048/561837) and with either BD Horizon™ BV480 Rat IgG2b, κ Isotype Control (Cat. No. 565649; Left Plots) or BD Horizon™ BV480 Rat Anti-Mouse CD274 (PD-L1) antibody (Cat. No. 568589/568590; Right Plots) at 1.0 µg/test. DAPI (4',6-Diamidino-2-Phenylindole, Dihydrochloride) Solution (Cat. No. 564907) was added to cells right before analysis. Bivariate pseudocolor density plots showing the correlated expression of CD274 (PD-L1) [or Ig Isotype control staining] versus CD4 were derived from gated events with the forward and side light scatter characteristics of viable (DAPI-negative) leucocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 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™
Cd274; Pdl1; Pdcd1l1; Pdcd1lg1; B7 homolog 1; B7h1
Mouse (QC Testing)
Rat LEW, also known as Lewis IgG2b, κ
Mouse PD-L1
Flow cytometry (Routinely Tested)
0.2 mg/ml
60533
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. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  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. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
Antibody Details
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10F.9G2

The 10F.9G2 monoclonal antibody specifically recognizes Programmed cell death 1 ligand 1 (PDCD1 ligand 1) which is also known as Programmed death ligand 1 (PD-L1), B7 homolog 1 (B7-H1), or CD274. CD274 (PD-L1) is a 43-kDa type I transmembrane glycoprotein that is encoded by Cd274 which belongs to the B7 family within the Ig superfamily. CD274 (PD-L1) is expressed at low levels on resting peripheral T and B lymphocytes, monocytes, macrophages, dendritic cells (DC), and NK cells and undergoes upregulated expression upon cellular activation. PD-L1 (CD274) is also widely expressed on nonhematopoietic cell types including epithelial cells, endothelial cells, placental trophoblasts, and tumor cells. CD274 (PD-L1) and CD273 (PD-L2) serve as ligands for the CD279 (Programmed cell death protein 1/PD-1) inhibitory receptor. CD274 (PD-L1)-mediated signaling through CD279 (PD-1) regulates T cell responses in lymphoid and nonlymphoid tissues that are important for ensuring protective immunity while maintaining peripheral tolerance. This immune signaling checkpoint may suppress antitumor immune responses and prevent tumor rejection. The 10F.9G2 antibody reportedly blocks CD274 (PD-L1) binding to CD279 (PD-1) and can enhance the proliferation and effector responses of activated T cells including cytokine production and cytolytic activity.

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
Citations & References
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View product citations for antibody "568590" on CiteAb

Development References (6)

  1. Eppihimer MJ, Gunn J, Freeman GJ, et al. Expression and regulation of the PD-L1 immunoinhibitory molecule on microvascular endothelial cells. Microcirculation. 2002; 9(2):133-145. (Immunogen: ELISA, Flow cytometry, Immunohistochemistry, Radioimmunoassay). View Reference
  2. Freeman GJ, Long AJ, Iwai Y, et al. Engagement of PD-1 immunoinhibitory receptor by a novel B7 family member leads to negative regulation of lymphocyte activation. J Exp Med. 2000; 192:1027-1034. (Biology). View Reference
  3. Iraolagoitia XL, Spallanzani RG, Torres NI, et al. NK Cells Restrain Spontaneous Antitumor CD8+ T Cell Priming through PD-1/PD-L1 Interactions with Dendritic Cells.. J Immunol. 2016; 197(3):953-61. (Clone-specific: Flow cytometry). View Reference
  4. Paterson AM, Brown KE, Keir ME, et al. The programmed death-1 ligand 1:B7-1 pathway restrains diabetogenic effector T cells in vivo.. J Immunol. 2011; 187(3):1097-105. (Clone-specific: Blocking). View Reference
  5. 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
  6. Sharpe AH, Wherry EJ, Ahmed R, Freeman GJ. The function of programmed cell death 1 and its ligands in regulating autoimmunity and infection.. Nat Immunol. 2007; 8(3):239-45. (Biology). View Reference
View All (6) View Less

 

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