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PE-CF594 Mouse Anti-Human PD-L1 (CD274)

BD Horizon™ PE-CF594 Mouse Anti-Human PD-L1 (CD274)

Clone IPI652.rMAb (also known as IPI652.rMAb)

(RUO)
PE-CF594 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™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; dashed line histogram) or BD Horizon™ PE-CF594 Mouse Anti-Human PD-L1 (CD274) antibody (Cat. No. 568984/568985; 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-L1 (CD274) expression [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™ PE-CF594 Mouse IgG1, κ Isotype Control (Cat. No. 562292; dashed line histogram) or BD Horizon™ PE-CF594 Mouse Anti-Human PD-L1 (CD274) antibody (Cat. No. 568984/568985; 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-L1 (CD274) expression [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.
Product Details
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BD Horizon™
B7H1; PD-L1; PDCD1L1; PDCD1LG1; PDL1; hPD-L1
Human (QC Testing)
Mouse IgG1, κ
Human PD-L1
Flow cytometry (Routinely Tested)
5 µl
29126
Aqueous buffered solution containing BSA and ≤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.

Product Notices

  1. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  2. 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.
  3. 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.
  4. 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).
  5. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  6. An isotype control should be used at the same concentration as the antibody of interest.
  7. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  8. CF™ is a trademark of Biotium, Inc.
  9. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  10. This product is provided under an Agreement between BIOTIUM and BD Biosciences. The manufacture, use, sale, offer for sale, or import of this product is subject to one or more patents or pending applications owned or licensed by Biotium, Inc. This product, and only in the amount purchased by buyer, may be used solely for buyer’s own internal research, in a manner consistent with the accompanying product literature. No other right to use, sell or otherwise transfer (a) this product, or (b) its components is hereby granted expressly, by implication or by estoppel. This product is for research use only. Diagnostic uses require a separate license from Biotium, Inc. For information on purchasing a license to this product including for purposes other than research, contact Biotium, Inc., 3159 Corporate Place, Hayward, CA 94545, Tel: (510) 265-1027. Fax: (510) 265-1352. Email: btinfo@biotium.com.
  11. Because of the broad absorption spectrum of the tandem fluorochrome, extra care must be taken when using multi-laser cytometers, which may directly excite both PE and CF™594.
  12. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  13. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
568985 Rev. 1
Antibody Details
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IPI652.rMAb

The IPI652.rMAb recombinant 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.

568985 Rev. 1
Format Details
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PE-CF594
BD Horizon™ PE-CF594 dye is a part of the BD PE family of dyes. This tandem fluorochrome is comprised of a R-Phycoerythrin (PE) donor that has excitation maxima (Ex Max) of 496-nm and 566-nm and an acceptor dye with an emission maximum (Em Max) at 615-nm. PE-CF594, driven by BD innovation, 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 615 nm (e.g., a 610/20-nm bandpass filter). The donor dye can be excited by the Blue (488-nm), Green (532-nm) and yellow-green (561-nm) lasers and the acceptor dye can be excited by the green (532-nm) laser resulting in cross-laser excitation and fluorescence spillover. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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PE-CF594
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
615 nm
568985 Rev.1
Citations & References
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View product citations for antibody "568985" on CiteAb

Development References (6)

  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. (Biology). View Reference
  2. Burr ML, Sparbier CE, Chan YC, et al. CMTM6 maintains the expression of PD-L1 and regulates anti-tumour immunity.. Nature. 2017; 549(7670):101-105. (Biology). View Reference
  3. Dorfman DM, Brown JA, Shahsafaei A, Freeman GJ. Programmed death-1 (PD-1) is a marker of germinal center-associated T cells and angioimmunoblastic T-cell lymphoma. Am J Surg Pathol. 2006; 30:802-810. (Biology). 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. (Biology). 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. (Biology). View Reference
  6. 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. (Biology). View Reference
View All (6) View Less
568985 Rev. 1

 

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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.