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

Flow cytometric analysis of PD-1 (CD279) expression on human peripheral blood leucocyte populations. Human whole blood was stained with APC Mouse Anti-Human CD3 antibody (Cat No. 561811/555335/561810) and either PE Mouse IgG1, κ Isotype Control (Cat No. 554680; Left Plots) or PE Mouse Anti-Human PD-1 (CD279) antibody (Cat No. 567618/567617; Right Plots). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat No. 349202). Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Top Plots: Bivariate pseudocolor density plots showing correlated expression of PD-1 (CD279) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes.

Bottom Plots: Bivariate pseudocolor density plots showing correlated expression of PD-1 (CD279) [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 of PD-1 (CD279) expression on human peripheral blood leucocyte populations. Human whole blood was stained with APC Mouse Anti-Human CD3 antibody (Cat No. 561811/555335/561810) and either PE Mouse IgG1, κ Isotype Control (Cat No. 554680; Left Plots) or PE Mouse Anti-Human PD-1 (CD279) antibody (Cat No. 567618/567617; Right Plots). Erythrocytes were lysed with BD FACS™ Lysing Solution (Cat No. 349202). Flow cytometric analysis was performed using a BD LSRFortessa™ Flow Cytometer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.

Top Plots: Bivariate pseudocolor density plots showing correlated expression of PD-1 (CD279) [or Ig Isotype control staining] versus side light-scatter (SSC-A) signals were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes.

Bottom Plots: Bivariate pseudocolor density plots showing correlated expression of PD-1 (CD279) [or Ig Isotype control staining] versus CD3 were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes.

Product Details
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BD Pharmingen™
PD1; PDC1; PDCD1; Programmed cell death 1; SLEB2; hPD-1
Human (QC Testing)
Mouse IgG1, κ
Not Reported
Flow cytometry (Tested During Development), Immunofluorescence, Immunohistochemistry (Reported)
5 µl
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 unconjugated antibody and free dye were 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. 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. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
  6. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
  7. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
567618 Rev. 2
Antibody Details
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NAT105

The NAT105 monoclonal antibody specifically recognizes CD279, which is also known as Programmed cell death 1 (PD-1). CD279 is an ~55 kDa type I transmembrane glycoprotein in the CD28/CTLA-4 family within the Ig superfamily and is encoded by the Pdcd1 gene. CD279 has an extracellular region with an IgV-like domain and an intracellular region with an immunoreceptor tyrosine-based inhibitory motif (ITIM) and an immunoreceptor tyrosine-based switch motif (ITSM). CD279 is a suppressive immunoregulatory receptor expressed on CD4-CD8- thymocytes, activated T cells, B cells and myeloid cells. CD273 (also known as PD-L2 or B7-H1) and CD274 (also known as PD-L1 or B7-DC), are ligands of CD279 and members of the B7 gene family. Upon binding, CD279 inhibits 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 has been reported to be suitable for immunohistochemistry.

567618 Rev. 2
Format Details
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PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
altImg
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
567618 Rev.2
Citations & References
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Development References (13)

  1. Bekerman E, Hesselgesser J, Carr B, et al. PD-1 Blockade and TLR7 Activation Lack Therapeutic Benefit in Chronic Simian Immunodeficiency Virus-Infected Macaques on Antiretroviral Therapy.. Antimicrob Agents Chemother. 2019; 63(11):e01163-19. (Clone-specific: Flow cytometry). View Reference
  2. 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
  3. 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
  4. 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. (Biology). View Reference
  5. 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
  6. 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
  7. Kim KS, Sekar RR, Patil D, et al. Evaluation of programmed cell death protein 1 (PD-1) expression as a prognostic biomarker in patients with clear cell renal cell carcinoma.. Oncoimmunology. 7(4):e1413519. (Clone-specific: Immunohistochemistry). View Reference
  8. 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
  9. Linedale R, Schmidt C, King BT, et al. Elevated frequencies of CD8 T cells expressing PD-1, CTLA-4 and Tim-3 within tumour from perineural squamous cell carcinoma patients.. PLoS One. 2017; 12(4):e0175755. (Clone-specific: Immunohistochemistry). View Reference
  10. 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
  11. Pauken KE, Wherry EJ. Overcoming T cell exhaustion in infection and cancer. Trends Immunol. 2015; 36(4):265-273. (Biology). View Reference
  12. Roncador G, García Verdes-Montenegro JF, Tedoldi S, et al. Expression of two markers of germinal center T cells (SAP and PD-1) in angioimmunoblastic T-cell lymphoma. Haematologica. 2007; 92(8):1059-66. (Clone-specific: Immunofluorescence, Immunohistochemistry, Western blot). View Reference
  13. 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. (Biology). View Reference
View All (13) View Less
567618 Rev. 2

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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.