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Alexa Fluor™ 488 Mouse Anti-Human CD279 (PD-1)
Alexa Fluor™ 488 Mouse Anti-Human CD279 (PD-1)

Two-color flow cytometric analysis of CD279 expression on human peripheral blood lymphocytes. Human whole blood was stained with BD Horizon™ BUV395 Mouse Anti-Human CD3 antibody (Cat. No. 563546/ 563548) and either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human CD279 (PD-1) antibody (Cat. No. 567397/567398; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The two-color pseudocolor density plots show the correlated expression of CD279 (or Ig Isotype control staining) versus CD3 for gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software.

Two-color flow cytometric analysis of CD279 expression on human peripheral blood lymphocytes. Human whole blood was stained with BD Horizon™ BUV395 Mouse Anti-Human CD3 antibody (Cat. No. 563546/ 563548) and either Alexa Fluor™ 488 Mouse IgG1, κ Isotype Control (Cat. No. 565572; Left Plot) or Alexa Fluor™ 488 Mouse Anti-Human CD279 (PD-1) antibody (Cat. No. 567397/567398; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The two-color pseudocolor density plots show the correlated expression of CD279 (or Ig Isotype control staining) versus CD3 for gated events with the forward and side light-scatter characteristics of intact 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 Pharmingen™
PD1; hPD-1; hPD-l; PDCD1; PDC1; Programmed cell death 1; SLEB2; hSLE1
Human (QC Testing)
Mouse IgG1, κ
Human PD-1 Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl
5133
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 unreacted dye was 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. 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. 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. Source of all serum proteins is from USDA inspected abattoirs located in the United States.
  5. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  6. Alexa Fluor® 488 fluorochrome emission is collected at the same instrument settings as for fluorescein isothiocyanate (FITC).
  7. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  8. 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.
  9. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  10. Alexa Fluor™ is a trademark of Life Technologies Corporation.
567397 Rev. 1
Antibody Details
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EH12.1

The EH12.1 monoclonal antibody specifically binds to CD279 which is also known as Programmed cell death 1 (PD1). CD279 is an immunoregulatory receptor  expressed on activated T cells, B cells, and myeloid cells. It contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) in the cytoplasmic region. Mice deficient in CD279 show a breakdown of peripheral tolerance and manifest multiple autoimmune symptoms. PD-L1 and PD-L2 are ligands of CD279 and members of the B7 gene family. CD279:PD-Ligands interaction inhibits T cell proliferation and cytokine secretion. Reports suggest that the B7/CTLA-4 pathway primarily attenuates, limits, and/or terminates naïve T-cell activation in secondary lymphoid organs. The PD-ligand:CD279 pathway, on the other hand, may primarily attenuate, limit, and/or terminate T-, B-, and myeloid cell activation/effector function at sites of inflammation in the periphery.

567397 Rev. 1
Format Details
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Alexa Fluor™ 488
Alexa Fluor™ 488 Dye is part of the BD blue family of dyes. This is a small organic fluorochrome with an excitation maximum (Ex Max) at 494-nm and an emission maximum (Em Max) at 517-nm. Alexa Fluor™ 488 is designed to be excited by the Blue laser (488 nm) and detected using an optical filter centered near 520-nm (e.g., a 530/30-nm bandpass filter). Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
Alexa Fluor™ 488
Blue 488 nm
494 nm
517 nm
567397 Rev.1
Citations & References
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Development References (9)

  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. 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. (Immunogen: Flow cytometry, Immunohistochemistry). View Reference
  4. 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
  5. Kanai T, Totsuka T, Uraushihara K, et al. Blockade of B7-H1 suppresses the development of chronic intestinal inflammation. J Immunol. 2003; 171(8):4156-4163. (Biology). View Reference
  6. 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
  7. 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. (Immunogen: Immunoprecipitation). View Reference
  8. Van der Sluis RM, Kumar NA, Pascoe RD, et al. Combination Immune Checkpoint Blockade to Reverse HIV Latency. J Immunol. 2020; 204:1-13. (Clone-specific: Flow cytometry, Fluorescence activated cell sorting). View Reference
  9. Velu V, Kannanganat S, Ibegbu C, et al. Elevated expression levels of inhibitory receptor programmed death 1 on simian immunodeficiency virus-specific CD8 T cells during chronic infection but not after vaccination. J Virol. 2007; 81(11):5819-5828. (Clone-specific: Blocking, Flow cytometry, Functional assay, Inhibition). View Reference
View All (9) View Less
567397 Rev. 1

Please refer to Support Documents for Quality Certificates


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.

Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.