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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.
BD Horizon™ PE-CF594 Mouse Anti-Human CD279 (PD-1)
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Preparation And Storage
Product Notices
- 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).
- An isotype control should be used at the same concentration as the antibody of interest.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- 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.
- When excited by the yellow-green (561-nm) laser, the fluorescence may be brighter than when excited by the blue (488-nm) laser.
- 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.
- 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.
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Companion Products
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).
Development References (8)
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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
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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
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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
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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
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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
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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
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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
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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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For Research Use Only. Not for use in diagnostic or therapeutic procedures.