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PE-CF594 Mouse Anti-Human CD279 (PD-1)
PE-CF594 Mouse Anti-Human CD279 (PD-1)
Flow cytometric analysis of CD279 (PD-1) expression by resting and activated human leucocytes.      Panels 1 and 2. CD279 (PD-1) expression on resting peripheral blood leucocytes. Fresh whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426/562427) and either BD Horizon PE-CF594 Mouse IgG1 κ Isotype Control (Cat. No. 562292; Left Plots) or BD Horizon PE-CF594 Mouse Anti-Human CD279 (PD-1) antibody (Cat. No. 566845/566846; Right Plots). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Bivariate pseudocolor density plots showing the correlated expression of CD279 (PD-1) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals [Panel 1] or CD3 [Panel 2] were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations.      Panel 3. CD279 (PD-1) expression on stimulated peripheral blood lymphocytes. Phytohemagglutinin-stimulated (3 days) peripheral blood mononuclear cells were stained with either BD Horizon PE-CF594 Mouse IgG1 κ Isotype Control (dashed line histogram) or BD Horizon PE-CF594 Mouse Anti-Human CD279 (PD-1) antibody (solid line histogram). The fluorescence histogram showing CD279 (PD-1) expression (or Ig Isotype control staining) was derived from gated events forward and side light-scatter characteristics of viable lymphoblasts.      Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.
Flow cytometric analysis of CD279 (PD-1) expression by resting and activated human leucocytes.      Panels 1 and 2. CD279 (PD-1) expression on resting peripheral blood leucocytes. Fresh whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 562426/562427) and either BD Horizon PE-CF594 Mouse IgG1 κ Isotype Control (Cat. No. 562292; Left Plots) or BD Horizon PE-CF594 Mouse Anti-Human CD279 (PD-1) antibody (Cat. No. 566845/566846; Right Plots). Erythrocytes were lysed with BD FACS Lysing Solution (Cat. No. 349202). Bivariate pseudocolor density plots showing the correlated expression of CD279 (PD-1) [or Ig Isotype control staining] versus side-light scatter (SSC-A) signals [Panel 1] or CD3 [Panel 2] were derived from gated events with the forward and side-light scatter characteristics of intact leucocyte populations.      Panel 3. CD279 (PD-1) expression on stimulated peripheral blood lymphocytes. Phytohemagglutinin-stimulated (3 days) peripheral blood mononuclear cells were stained with either BD Horizon PE-CF594 Mouse IgG1 κ Isotype Control (dashed line histogram) or BD Horizon PE-CF594 Mouse Anti-Human CD279 (PD-1) antibody (solid line histogram). The fluorescence histogram showing CD279 (PD-1) expression (or Ig Isotype control staining) was derived from gated events forward and side light-scatter characteristics of viable lymphoblasts.      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™
PD1; hPD-1; hPD-l; PDCD1; PDC1; Programmed cell death 1; SLEB2
Human (QC Testing)
Mouse IgG1, κ
Human PD-1 Transfected Cell Line
Flow cytometry (Routinely Tested)
5 µl
IX 37
5133
AB_2869903
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 with BD Horizon™ PE-CF594 under optimum conditions, and unconjugated antibody and free PE-CF594 were removed.

Product Notices

  1. 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).
  2. An isotype control should be used at the same concentration as the antibody of interest.
  3. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  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. When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
  6. 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.
  7. 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.
  8. CF™ is a trademark of Biotium, Inc.
  9. 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.
  10. Texas Red is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  11. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  12. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  13. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Antibody Details
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MIH4

The MIH4 monoclonal antibody specifically binds to CD279, which is also known as, Programmed cell death 1 (PD-1). CD279 is a type I transmembrane glycoprotein that belongs to the Ig superfamily. CD279 is an immunoregulatory receptor that is expressed on expressed on subsets of thymocytes, activated T cells, B cells and myeloid cells. CD279 contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) in its cytoplasmic region. CD273 (PD-L2) and CD274 (PD-L1) are ligands of CD279 and are members of the B7 gene family. Interaction of CD279 with its ligands results in inhibition of T cell proliferation and cytokine secretion. CD279 may play roles in supporting self-tolerance, reducing autoimmunity, or promoting T cell exhaustion associated with certain diseases.

This antibody is conjugated to BD Horizon PE-CF594, which has been developed exclusively by BD Biosciences as a better alternative to PE-Texas Red®. PE-CF594 excites and emits at similar wavelengths to PE-Texas Red® yet exhibits improved brightness and spectral characteristics. Due to PE having maximal absorption peaks at 496 nm and 564 nm, PE-CF594 can be excited by the blue (488-nm), green (532-nm) and yellow-green (561-nm) lasers and can be detected with the same filter set as PE-Texas Red® (eg, 610/20-nm filter).

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
Citations & References
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View product citations for antibody "566845" on CiteAb

Development References (8)

  1. Bennett F, Luxenberg D, Ling V, et al. Program death-1 engagement upon TCR activation has distinct effects on costimulation and cytokine-driven proliferation: attenuation of ICOS, IL-4, and IL-21, but not CD28, IL-7, and IL-15 responses. J Immunol. 2003; 170(2):711-718. (Biology). View Reference
  2. Carter L, Fouser LA, Jussif J, et al. PD-1:PD-L inhibitory pathway affects both CD4(+) and CD8(+) T cells and is overcome by IL-2. Eur J Immunol. 2002; 32:634-643. (Biology). View Reference
  3. 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
  4. Igarashi H, Cao Y, Iwai H, et al. GITR ligand-costimulation activates effector and regulatory functions of CD4+ T cells. Biochem Biophys Res Commun. 2008; 369(4):1134-1138. (Clone-specific: Fluorescence microscopy, Immunofluorescence, Immunohistochemistry). 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. Nishimura H, Minato N, Nakano T, Honjo T. Immunological studies on PD-1 deficient mice: implication of PD-1 as a negative regulator for B cell responses. Int Immunol. 1998; 10(10):1563-1572. (Biology). View Reference
  7. Youngnak P, Kozono Y, Kozono H, et al. Differential binding properties of B7-H1 and B7-DC to programmed death-1. Biochem Biophys Res Commun. 2003; 307(3):672-677. (Immunogen: Flow cytometry). View Reference
  8. van Dongen JJ, Lhermitte L, Böttcher S, et al. EuroFlow antibody panels for standardized n-dimensional flow cytometric immunophenotyping of normal, reactive and malignant leukocytes. Leukemia. 2012; 26(9):1908-1975. (Clone-specific: Flow cytometry). View Reference
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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.