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Alexa Fluor® 647 Mouse Anti-Human CD279 (PD-1)
Alexa Fluor® 647 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 Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 565571; Left Plots) or Alexa Fluor® 647 Mouse Anti-Human CD279 (PD-1) antibody (Cat. No. 566850/566851; 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 Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (dashed line histogram) or Alexa Fluor® 647 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 Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (Cat. No. 565571; Left Plots) or Alexa Fluor® 647 Mouse Anti-Human CD279 (PD-1) antibody (Cat. No. 566850/566851; 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 Alexa Fluor® 647 Mouse IgG1 κ Isotype Control (dashed line histogram) or Alexa Fluor® 647 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 Pharmingen™
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_2869905
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 Alexa Fluor® 647 under optimum conditions, and unreacted Alexa Fluor® 647 was 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. The Alexa Fluor®, Pacific Blue™, and Cascade Blue® dye antibody conjugates in this product are sold under license from Molecular Probes, Inc. for research use only, excluding use in combination with microarrays, or as analyte specific reagents. The Alexa Fluor® dyes (except for Alexa Fluor® 430), Pacific Blue™ dye, and Cascade Blue® dye are covered by pending and issued patents.
  6. Alexa Fluor® is a registered trademark of Molecular Probes, Inc., Eugene, OR.
  7. Alexa Fluor® 647 fluorochrome emission is collected at the same instrument settings as for allophycocyanin (APC).
  8. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  9. This product is provided under an intellectual property license between Life Technologies Corporation and BD Businesses. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The buyer cannot sell or otherwise transfer (a) this product (b) its components or (c) materials made using this product or its components to a third party or otherwise use this product or its components or materials made using this product or its components for Commercial Purposes. Commercial Purposes means any activity by a party for consideration and may include, but is not limited to: (1) use of the product or its components in manufacturing; (2) use of the product or its components to provide a service, information, or data; (3) use of the product or its components for therapeutic, diagnostic or prophylactic purposes; or (4) resale of the product or its components, whether or not such product or its components are resold for use in research. For information on purchasing a license to this product for any other use, contact Life Technologies Corporation, Cell Analysis Business Unit Business Development, 29851 Willow Creek Road, Eugene, OR 97402, USA, Tel: (541) 465-8300. Fax: (541) 335-0504.
  10. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  11. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
566851 Rev. 1
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.

        

566851 Rev. 1
Format Details
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Alexa Fluor™ 647
Alexa Fluor™ 647 Dye is part of the BD red family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 653-nm and an emission maximum (Em Max) at 669-nm. Alexa Fluor 647 is designed to be excited by the Red laser (627-640 nm) and detected using an optical filter centered near 520-nm (e.g., a 660/20 nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 647
Red 627-640 nm
653 nm
669 nm
566851 Rev.1
Citations & References
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View product citations for antibody "566851" 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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566851 Rev. 1

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For Research Use Only. Not for use in diagnostic or therapeutic procedures.