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R718 Mouse Anti-Human CD279 (PD-1)
R718 Mouse Anti-Human CD279 (PD-1)

Two-color flow cytometric analysis of CD279 (PD-1) expression on human peripheral blood lymphocytes. Human whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 563797/563798) and either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plot) or BD Horizon™ R718 Mouse Anti-Human CD279 (PD-1) antibody (Cat. No. 566974/566975; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The two-color flow cytometric contour 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 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 (PD-1) expression on human peripheral blood lymphocytes. Human whole blood was stained with BD Horizon™ BV421 Mouse Anti-Human CD3 antibody (Cat. No. 563797/563798) and either BD Horizon™ R718 Mouse IgG1, κ Isotype Control (Cat. No. 566928; Left Plot) or BD Horizon™ R718 Mouse Anti-Human CD279 (PD-1) antibody (Cat. No. 566974/566975; Right Plot). The erythrocytes were lysed with BD FACS™ Lysing Solution (Cat. No. 349202). The two-color flow cytometric contour 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 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; hSLE1
Human (QC Testing)
Mouse IgG1, κ
Human PD-1 Recombinant Protein
Flow cytometry (Routinely Tested)
5 µl
5133
AB_2869981
Aqueous buffered solution containing ≤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 CompBead 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. Alexa Fluor® is a registered trademark of Life Technologies Corporation.
  5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
  6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  7. This product is provided under an Agreement between BIOTIUM and BD Biosciences. 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.
566975 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.

The antibody was conjugated to BD Horizon Red 718, which has been developed exclusively for BD Biosciences as a better alternative to Alexa Fluor® 700. BD Horizon Red 718 can be excited by the red laser (628 – 640 nm) and, with an Em Max around 718 nm, it can be detected using a 730/45 nm filter. Due to similar excitation and emission properties, we do not recommend using R718 in combination with APC-R700 or Alexa Fluor® 700.

566975 Rev. 1
Format Details
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R718
The BD Horizon™ Red 718 (R718) Dye is part of the BD red family of dyes. It is a small organic fluorochrome with an excitation maximum (Ex Max) at 695-nm and an emission maximum (Em Max) at 718-nm. Driven by BD innovation, R718 is designed to be excited by the red laser (627–640-nm) and detected using an optical filter centered near 720-nm (e.g., a 720/40-nm bandpass filter). R718 is a brighter alternative to Alexa Fluor™ 700. R718 is also a bright small molecule alternative to APC-R700 with lower spread into the APC detector. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
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R718
Red 627-640 nm
695 nm
718 nm
566975 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
566975 Rev. 1

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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.