Skip to main content Skip to navigation
PE Rat Anti-Mouse CD274 (PD-L1)
PE Rat Anti-Mouse CD274 (PD-L1)

Multicolor flow cytometric analysis of CD274 (PD-L1) expression on resting and activated mouse splenocytes. C57BL/6 mouse splenic leucocytes were either not activated (Upper Plots) or activated (Lower Plots) by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3 antibody (Cat. No. 553057) 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 APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051) and with either PE Rat IgG2b, κ Isotype Control (Cat. No. 555848; Left Plots) or PE Rat Anti-Mouse CD274 (PD-L1) antibody (Cat. No. 568085; Right Plots) at 0.25 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 FACSCelesta™ 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. C57BL/6 mouse splenic leucocytes were either not activated (Upper Plots) or activated (Lower Plots) by culture with plate-bound Purified NA/LE Hamster Anti-Mouse CD3 antibody (Cat. No. 553057) 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 APC Rat Anti-Mouse CD4 antibody (Cat. No. 553051) and with either PE Rat IgG2b, κ Isotype Control (Cat. No. 555848; Left Plots) or PE Rat Anti-Mouse CD274 (PD-L1) antibody (Cat. No. 568085; Right Plots) at 0.25 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 FACSCelesta™ Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

商品详情
Down Arrow Up Arrow


BD Pharmingen™
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
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO


准备和存储

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.

推荐的实验流程

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 CompBead to ensure that BD CompBeads are appropriate for your specific cellular application.

商品通知

  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. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  4. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  5. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
  6. An isotype control should be used at the same concentration as the antibody of interest.
568085 Rev. 1
抗体详情
Down Arrow Up Arrow
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.

568085 Rev. 1
格式详情
Down Arrow Up Arrow
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
568085 Rev.1
报价单和参考
Down Arrow Up Arrow

研发参考 (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). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
  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). 查看参考
查看所有文件 (6) 查看更少内容
568085 Rev. 1

Please refer to Support Documents for Quality Certificates


Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described


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.