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Two-color flow cytometric analysis of CD279 (PD-1) expression on human peripheral blood lymphocytes. Human whole blood was stained with PE Mouse Anti-Human CD3 antibody (Cat. No. 555333/561808/561809) and either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (Cat. No. 562438; Left Panel) or BD Horizon BV421 Mouse Anti-Human CD279 (PD-1) antibody (Cat. No. 564323; Right Panel). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). Two-color flow cytometric dot plots showing the correlated expression of CD279 (PD-1) [or Ig Isotype control staining] versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.

Flow cytometric analysis of CD279 (PD-1) expression on stimulated human peripheral blood lymphocytes. Phytohemagglutinin-stimulated (3 days) peripheral blood mononuclear cells were stained with either BD Horizon™ BV421 Mouse IgG1, κ Isotype Control (dashed line histogram) or BD Horizon BV421 Mouse Anti-Human CD279 (PD-1) antibody (solid line histogram). The fluorescence histograms were derived from events with the forward and side light-scatter characteristics of viable lymphoblasts. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.


BD Horizon™ BV421 Mouse Anti-Human CD279 (PD-1)

BD Horizon™ BV421 Mouse Anti-Human CD279 (PD-1)

Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
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.
- 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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Pacific Blue™ is a trademark of Molecular Probes, Inc., Eugene, OR.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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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.
The antibody was conjugated to BD Horizon™ BV421 which is part of the BD Horizon Brilliant™ Violet family of dyes. With an Ex Max of 407-nm and Em Max at 421-nm, BD Horizon BV421 can be excited by the violet laser and detected in the standard Pacific Blue™ filter set (eg, 450/50-nm filter). BD Horizon BV421 conjugates are very bright, often exhibiting a 10 fold improvement in brightness compared to Pacific Blue conjugates.

Development References (6)
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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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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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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.