PD-1 Antibody [Unconjugated]

Western blot shows lysate of human thymus tissue. PVDF membrane was probed with 2 µg/mL of Goat Anti-Human PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1086) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for PD‑1 at approximately 40-50 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Western blot shows lysates of HEK293 human embryonic kidney cell line either mock transfected or transfected with human PD-1. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1086) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). Specific bands were detected for PD‑1 at approximately 40-80 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

PD‑1 was detected in immersion fixed paraffin-embedded sections of human lymph node using Goat Anti-Human PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1086) at 3 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Human peripheral blood mononuclear cells (PBMCs) either (A) untreated or (B) treated with 5 μg/mL PHA overnight were stained with Goat Anti-Human PD‑1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1086) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107) and Mouse Anti-Human CD3 epsilon APC‑conjugated Monoclonal Antibody (Catalog # FAB100A). Quadrant markers were set based on control antibody staining (Catalog # F0107). View our protocol for Staining Membrane-associated Proteins.

• Reactivity: Hu
• Applications: WB, Flow, IHC, B/N, CyTOF-ready, ELISA(Cap)
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

PD-1 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human PD-1
  Leu25-Gln167
  Accession # Q8IX89
• Specificity: Detects human PD-1 in ELISAs and Western blots. In sandwich ELISAs, less than 2% cross-reactivity with recombinant mouse PD‑1 and less than 0.2% cross-reactivity with recombinant human (rh) CD28, rhICOS, and rhCTLA‑4 is observed.
• Details of Functionality: ActivityAssaywCitation not found None None None None None
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: PDCD1
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Western Blot 0.5-2 ug/mL
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 5-15 ug/mL
  • Blockade of Receptor-ligand Interaction
  • CyTOF-ready
  • ELISA Capture (Matched Antibody Pair) 0.2-0.8 ug/mL
  • ELISA Detection (Matched Antibody Pair) 0.1-0.4 ug/mL
  • ELISA Standard (Matched Pair)
• Application Notes: ELISA Capture: Human PD-1 Antibody (Catalog # AF1086)
  ELISA Detection: Human PD-1 Biotinylated Antibody (Catalog # BAF1086)
  Standard: Recombinant Human PD-1 Fc Chimera (Catalog # 1086-PD)
• Control:

  PD-1 Knockout HeLa Cell Lysate

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for PD-1 Antibody [Unconjugated]

• CD279 antigen
• CD279
• hPD-1
• PD1
• PD-1
• PD1hPD-l
• PDCD1
• programmed cell death 1
• programmed cell death protein 1
• Protein PD-1
• SLEB2

Background

Programmed Death-1 (PD-1) is a type I transmembrane protein belonging to the CD28/CTLA-4 family of immunoreceptors that mediate signals for regulating immune responses (1). Members of the CD28/CTLA-4 family have been shown to either promote T cell activation (CD28 and ICOS) or down-regulate T cell activation (CTLA-4 and PD-1) (2). PD-1 is expressed on activated T cells, B cells, myeloid cells, and on a subset of thymocytes. In vitro, ligation of PD-1 inhibits TCR-mediated T-cell proliferation and production of IL-1, IL-4, IL-10, and IFN-gamma. In addition, PD-1 ligation also inhibits BCR mediated signaling. PD-1 deficient mice have a defect in peripheral tolerance and spontaneously develop autoimmune diseases (2, 3).

Two B7 family proteins, PD-L1 (also called B7-H1) and PD-L2 (also known as B7-DC), have been identified as PD-1 ligands. Unlike other B7 family proteins, both
PD‑L1 and PD-L2 are expressed in a wide variety of normal tissues including heart, placenta, and activated spleens (4). The wide expression of PD-L1 and PD-L2 and the inhibitor effects on PD-1 ligation indicate that PD-1 might be involved in the regulation of peripheral tolerance and may help prevent autoimmune diseases (2).

The human PD-1 gene encodes a 288 amino acid (aa) protein with a putative 20 aa signal peptide, a 148 aa extracellular region with one immunoglobulin-like V-type domain, a 24 aa transmembrane domain, and a 95 aa cytoplasmic region. The cytoplasmic tail contains two tyrosine residues that form the immuno-receptor
tyrosine-based inhibitory motif (ITIM) and immunoreceptor tyrosine-based switch motif (ITSM) that are important in mediating PD-1 signaling. Mouse and human PD-1 share approximately 60% aa sequence identity (4).

1. Ishida, Y. et al. (1992) EMBO J. 11:3887.
2. Nishimura, H. and T. Honjo (2001) Trends in Immunol. 22:265.
3. Latchman, Y. et al. (2001) Nature Immun. 2:261.
4. Carreno, B.M. and M. Collins (2002) Annu. Rev. Immunol. 20:29.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for PD-1 Antibody (AF1086)(23)

Publications using AF1086

• JE Eckel-Pass, TH Ho, DJ Serie, JC Cheville, R Houston Th, BA Costello, H Dong, ED Kwon, BC Leibovich, AS Parker Concordance of PD-1 and PD-L1 (B7-H1) in paired primary and metastatic clear cell renal cell carcinoma Cancer Med, 2019;0(0):. 2019 [PMID: 31829518] (IHC-P, Human)
• S Li, JM Folkvord, KJ Kovacs, RK Wagstaff, G Mwakalundw, AK Rendahl, EG Rakasz, E Connick, PJ Skinner Low levels of SIV-specific CD8+ T cells in germinal centers characterizes acute SIV infection PLoS Pathog., 2019;15(3):e1007311. 2019 [PMID: 30897187] (IHC, Primate - Macaca mulatta (Rhesus Macaque))
• B Li, TN Song, FR Wang, C Yin, Z Li, JP Lin, YQ Meng, HM Feng, T Jing Tumor-derived exosomal HMGB1 promotes esophageal squamous cell carcinoma progression through inducing PD1+ TAM expansion Oncogenesis, 2019;8(3):17. 2019 [PMID: 30796203] (Bioassay, Human)
• M Sampedro-N, A Serrano-So, M Adrados, JM Cameselle-, C Blanco-Car, JM Cabezas-Ag, R Martínez-H, E Martín-Pér, JL Muñoz de N, JÁ Díaz, R García-Cen, J Caneiro-Gó, I Abdulkader, R González-A, M Marazuela Analysis of expression of the PD-1/PD-L1 immune checkpoint system and its prognostic impact in gastroenteropancreatic neuroendocrine tumors Sci Rep, 2018;8(1):17812. 2018 [PMID: 30546030] (IHC-P, Human)
• F Wang, B Li, Y Wei, Y Zhao, L Wang, P Zhang, J Yang, W He, H Chen, Z Jiao, Y Li Tumor-derived exosomes induce PD1+ macrophage population in human gastric cancer that promotes disease progression Oncogenesis, 2018;7(5):41. 2018 [PMID: 29799520] (Functional Assay, Human)
• S Du, N McCall, K Park, Q Guan, P Fontina, A Ertel, T Zhan, AP Dicker, B Lu Blockade of Tumor-Expressed PD-1 promotes lung cancer growth Oncoimmunology, 2018;7(4):e1408747. 2018 [PMID: 29632720] (IHC-P, Human)
• W Fan, AJ Demers, Y Wan, Q Li Altered Ratio of T Follicular Helper Cells to T Follicular Regulatory Cells Correlates with Autoreactive Antibody Response in Simian Immunodeficiency Virus-Infected Rhesus Macaques J. Immunol., 2018;0(0):. 2018 [PMID: 29610141] (Flow Cytometry, Primate)
• MA Rahman, KM McKinnon, TS Karpova, DA Ball, DJ Venzon, W Fan, G Kang, Q Li, M Robert-Gur Associations of Simian Immunodeficiency Virus (SIV)-Specific Follicular CD8+T Cells with Other Follicular T Cells Suggest Complex Contributions to SIV Viremia Control J. Immunol., 2018;0(0):. 2018 [PMID: 29507105] (IHC, Primate)
• D Amodio, N Cotugno, G Macchiarul, S Rocca, Y Dimopoulos, MR Castrucci, R De Vito, FM Tucci, AB McDermott, S Narpala, P Rossi, RA Koup, P Palma, C Petrovas Quantitative Multiplexed Imaging Analysis Reveals a Strong Association between Immunogen-Specific B Cell Responses and Tonsillar Germinal Center Immune Dynamics in Children after Influenza Vaccination J. Immunol., 2017;0(0):. 2017 [PMID: 29237774] (IHC, Human)
• Y Liu, Y Cheng, Y Xu, Z Wang, X Du, C Li, J Peng, L Gao, X Liang, C Ma Increased expression of programmed cell death protein 1 on NK cells inhibits NK-cell-mediated anti-tumor function and indicates poor prognosis in digestive cancers Oncogene, 2017;0(0):. 2017 [PMID: 28692048] (Functional Assay, Human)
• ELV Silveira, KA Rogers, S Gumber, P Amancha, P Xiao, SM Woollard, SN Byrareddy, MM Teixeira, F Villinger Immune Cell Dynamics in Rhesus Macaques Infected with a Brazilian Strain of Zika Virus J. Immunol., 2017;0(0):. 2017 [PMID: 28667164] (IHC, Primate - Macaca mulatta (Rhesus Macaque))
• P Martinez-M, K Tran, J Guenaga, G Lindgren, M Àdori, Y Feng, GE Phad, N Vázquez Be, S Bale, J Ingale, V Dubrovskay, S O'Dell, L Pramanik, M Spångberg, M Corcoran, K Loré, JR Mascola, RT Wyatt, GB Karlsson H Particulate Array of Well-Ordered HIV Clade C Env Trimers Elicits Neutralizing Antibodies that Display a Unique V2 Cap Approach Immunity, 2017;46(5):804-817.e7. 2017 [PMID: 28514687] (IHC, Primate)
• Joseph M Obeid PD-L1, PD-L2 and PD-1 expression in metastatic melanoma: Correlation with tumor-infiltrating immune cells and clinical outcome Oncoimmunology, 2016;5(11):e1235107. 2016 [PMID: 27999753] (IHC-P, Human)
• PT Nghiem, S Bhatia, EJ Lipson, RR Kudchadkar, NJ Miller, L Annamalai, S Berry, EK Chartash, A Daud, SP Fling, PA Friedlande, HM Kluger, HE Kohrt, L Lundgren, K Margolin, A Mitchell, T Olencki, DM Pardoll, SA Reddy, EM Shantha, WH Sharfman, E Sharon, She PD-1 Blockade with Pembrolizumab in Advanced Merkel-Cell Carcinoma N Engl J Med, 2016;0(0):. 2016 [PMID: 27093365] (IHC-P, Human)
• Vargas-Inchaustegui D, Demers A, Shaw J, Kang G, Ball D, Tuero I, Musich T, Mohanram V, Demberg T, Karpova T, Li Q, Robert-Guroff M Vaccine Induction of Lymph Node-Resident Simian Immunodeficiency Virus Env-Specific T Follicular Helper Cells in Rhesus Macaques. J Immunol, 2016;196(4):1700-10. 2016 [PMID: 26773147] (IHC, Primate - Macaca mulatta (Rhesus Macaque))
• Zhang Z, Cheng L, Zhao J, Li G, Zhang L, Chen W, Nie W, Reszka-Blanco N, Wang F, Su L Plasmacytoid dendritic cells promote HIV-1-induced group 3 innate lymphoid cell depletion. J Clin Invest, 2015;125(9):3692-703. 2015 [PMID: 26301812] (IHC-P, Mouse)
• Melendreras S, Martinez-Camblor P, Menendez A, Bravo-Mendoza C, Gonzalez-Vidal A, Coto E, Diaz-Corte C, Ruiz-Ortega M, Lopez-Larrea C, Suarez-Alvarez B Soluble co-signaling molecules predict long-term graft outcome in kidney-transplanted patients. PLoS ONE, 2014;9(12):e113396. 2014 [PMID: 25478957] (ELISA Development, Human)
• Hong J, Amancha P, Rogers K, Courtney C, Havenar-Daughton C, Crotty S, Ansari A, Villinger F Early lymphoid responses and germinal center formation correlate with lower viral load set points and better prognosis of simian immunodeficiency virus infection. J Immunol, 2014;193(2):797-806. 2014 [PMID: 24907346] (IHC, Primate - Macaca mulatta (Rhesus Macaque))
• CD4 T follicular helper cell dynamics during SIV infection. J. Clin. Invest., 2012;122(9):3281-94. 2012 [PMID: 22922258] (IHC-P, Primate - Macaca mulatta (Rhesus Macaque))
• Schatton T, Schutte U, Frank NY, Zhan Q, Hoerning A, Robles SC, Zhou J, Hodi FS, Spagnoli GC, Murphy GF, Frank MH Modulation of T-cell activation by malignant melanoma initiating cells. Cancer Res., 2010;70(2):697-708. 2010 [PMID: 20068175] (IHC, Human)
• Estes JD, Gordon SN, Zeng M, Chahroudi AM, Dunham RM, Staprans SI, Reilly CS, Silvestri G, Haase AT Early resolution of acute immune activation and induction of PD-1 in SIV-infected sooty mangabeys distinguishes nonpathogenic from pathogenic infection in rhesus macaques. J. Immunol., 2008;180(10):6798-807. 2008 [PMID: 18453600] (IHC-P, Primate - Cercocebus torquatus (Sooty Mangabey))
• Onlamoon N, Rogers K, Mayne AE, Pattanapanyasat K, Mori K, Villinger F, Ansari AA Soluble PD-1 rescues the proliferative response of simian immunodeficiency virus-specific CD4 and CD8 T cells during chronic infection. Immunology, 2008;124(2):277-93. 2008 [PMID: 18266718] (Neutralization, Primate - Macaca mulatta (Rhesus Macaque))
• Wan B, Nie H, Liu A, Feng G, He D, Xu R, Zhang Q, Dong C, Zhang JZ Aberrant regulation of synovial T cell activation by soluble costimulatory molecules in rheumatoid arthritis. J. Immunol., 2006;177(12):8844-50. 2006 [PMID: 17142787] (ELISA Development, Human)

Review for PD-1 Antibody (AF1086) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Human.

reviewed by: Christopher England IF Human 07/09/2016

Summary

• Application: Immunofluorescence
• Sample Tested: Mouse salivary gland (one injected with human T cells and one with no human T cells)
• Species: Human
• Lot: ICA0714071

Blocking

• Blocking Details: 10% Normal Donkey Serum, 0.25% Triton-X in PBS, 1 hr at room temperature

Primary Anitbody

• Dilution Ratio: 1:200, 12 h, 4 degree Celcius, PBS w/0.25% Triton-X

Secondary Antibody

• Secondary Description: Donkey anti-Goat DyLight 488
• Secondary Manufacturer Cat#: NBP-1-72843
• Secondary Concentration: 1:4000

Details

• Detection Notes: Confocal microscopy, Nikon A1R with 488-nm laser
• Fixation Details: Frozen Sections, 4% Paraformaldehyde in PBS for 30 min at room temperature
• Wash Description: PBS, 15 min, 3-times after primary and 3-times after secondary

Comments

• Comments: The antibody worked well. Since goat-derived antibodies are not the best, we were happy to see that this one worked well.

Bioinformatics

• Gene Symbol: PDCD1
• Uniprot:
  • Q8IX89()