PD-L1 Antibody (130021) [Unconjugated]

Jurkat human acute T cell leukemia cell line was stained with Mouse Anti-Human PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB1561, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2Secondary Antibody (Catalog # F0102B).View our protocol for Staining Membrane-associated Proteins.

MDA-MB-231 human breast adenocarcinoma cell line was stained with Mouse Anti-Human PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB1561, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Phycoerythrin-conjugated Anti-Mouse IgG F(ab')2 Secondary Antibody (Catalog # F0102B). Adherent cells were prepared by either manual scraping or with TrypLE Express treatment with similar results. View our protocol for Staining Membrane-associated Proteins.

PD-L1/B7-H1 was detected in formalin fixed paraffin-embedded sections of human colon cancer using Mouse Anti-Human PD-L1/B7-H1 Monoclonal Antibody (Catalog # MAB1561) at 15 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was observed in the cytoplasm. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Formalin-fixed paraffin-embedded tissue sections of human colon cancer were probed for PDL1 mRNA (ACD RNAScope Probe, catalog # 600861; Fast Red chromogen, ACD catalog # 322360). Adjacent tissue section was processed for immunohistochemistry using mouse anti-human PDL1 monoclonal antibody (R&D Systems catalog # MAB1561) at 5ug/mL with overnight incubation at 4 degrees Celsius followed by incubation with anti-mouse IgG VisUCyte HRP Polymer Antibody (Catalog # VC001) and DAB chromogen (yellow-brown). Tissue was counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes.

Chemotherapeutics induce PD-L1 expression in NPC cells and PD-1 expression in NK cells via upregulation of NF-kappa B. NPC cells a or NK cells c were incubated with the NF-kappa B inhibitor BMS-345541 for 1 h before incubation with chemotherapeutics. NPC cells were transfected with NF-kappa B siRNA b or NK cells with NF-kappa B siRNA d for 16 h and then incubated with chemotherapeutics. Expression of NF-kappa B, PD-L1 and PD-1 was analyzed by immunoblot Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32737537), licensed under a CC-BY license. Not internally tested by R&D Systems.

Chemotherapeutics induce PD-L1 expression in NPC cells and PD-1 expression in NK cells via upregulation of NF-kappa B. NPC cells a or NK cells c were incubated with the NF-kappa B inhibitor BMS-345541 for 1 h before incubation with chemotherapeutics. NPC cells were transfected with NF-kappa B siRNA b or NK cells with NF-kappa B siRNA d for 16 h and then incubated with chemotherapeutics. Expression of NF-kappa B, PD-L1 and PD-1 was analyzed by immunoblot Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/32737537), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: Flow, IHC, CyTOF-ready, Dual ISH-IHC
• Clone: 130021
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

PD-L1 Antibody (130021) [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human PD-L1/B7-H1
  Phe19-Thr239
  Accession # Q9NZQ7
• Specificity: Detects human PD-L1/B7-H1 in direct ELISAs. In direct ELISAs, no cross-reactivity with recombinant human (rh) B7-1, -2, -H2, -H3, -H3b,
  -H4, rhPD-L2, recombinant mouse B7-H1, recombinant rat (rr) B7-1, or rrB7-2 is observed.
• Source: N/A
• Isotype: IgG1
• Clonality: Monoclonal
• Host: Mouse
• Gene: CD274
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • CyTOF-ready
  • Dual RNAscope ISH-IHC 3-25 ug/mL
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 8-25 ug/mL

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. See Certificate of Analysis for details.
  *Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Notes

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

Alternate Names for PD-L1 Antibody (130021) [Unconjugated]

• Avelumab
• B7-H
• B7H1
• B7-H1
• B7H1PDCD1L1
• CD274 antigenMGC142294
• CD274 molecule
• CD274
• PDCD1L1
• PDCD1LG1
• PDCD1LG1MGC142296
• PDL1
• PD-L1
• PD-L1B7 homolog 1
• PDL1PDCD1 ligand 1
• programmed cell death 1 ligand 1
• Programmed death ligand 1

Background

Human B7 homolog 1 (B7-H1), also called programmed death ligand 1 (PD-L1) and programmed cell death 1 ligand 1 (PDCD1L1), is a member of the growing B7 family of immune proteins that provide signals for both stimulating and inhibiting T cell activation. Other family members include B7-1, B7-2, B7-H2, PDL2 and B7-H3. B7 proteins are members of the immunoglobulin (Ig) superfamily. Their extracellular domains contain 2 Ig-like domains and all members have short cytoplasmic domains. Among the family members, there is about 20-25% amino acid identity. Human and mouse B7-H1 share approximately 70% amino acid sequence identity. B7-H1 has been identified as one of two ligands for programmed death-1 (PD-1), a member of the CD28 family of immunoreceptors. The B7-H1 gene encodes a 291 amino acid (aa) type I membrane precursor protein with a putative 18 aa signal peptide, a 220 aa extracellular domain, a 21 aa transmembrane region, and a 31 aa cytoplasmic domain. Human B7-H1 is constitutively expressed in several organs such as heart, skeletal muscle, placenta and lung, and in lower amounts in thymus, spleen, kidney and liver. B7-H1 expression is upregulated in a small fraction of activated T and B cells and a much larger fraction of activated monocytes. B7-H1 expression is also induced in dendritic cells and keratinocytes after IFN-gamma stimulation. Interaction of B7-H1 with PD-1 results in inhibition of TCR-mediated proliferation and cytokine production. The B7-H1:PD-1 pathway is involved in the negative regulation of some immune responses and may play an important role in the regulation of peripheral tolerance.

1. Nishimura, H. and T. Honjo (2001) Trends Immunol. 22:265.
2. Freeman, G.J. et al. (2000) J. Exp. Med. 192:1027.
3. Latchman, Y. et al. (2001) Nat. Immunol. 2:261.

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-L1 Antibody (MAB1561)(24)

Publications using MAB1561

• Q Niu, J Gao, K Zhao, X Chen, X Lin, C Huang, Y An, X Xiao, Q Wu, L Cui, P Zhang, L Wu, C Yang Fluid nanoporous microinterface enables multiscale-enhanced affinity interaction for tumor-derived extracellular vesicle detection Proceedings of the National Academy of Sciences of the United States of America, 2022-10-28;119(44):e2213236119. 10/28/2022 [PMID: 36306324]
• X Hou, X Shi, J Luo Efficacy and safety of camrelizumab (a PD-1 inhibitor) combined with chemotherapy as a neoadjuvant regimen in patients with locally advanced non-small cell lung cancer Oncology Letters, 2022-05-17;24(1):215. 2022-05-17 [PMID: 35720491] (IHC, Human)
• David Labib, Zhen Wang, Priya Prakash, Matthew Zimmer, Matthew D. Smith, Paul W. Frazel et al. Proteomic Alterations and Novel Markers of Neurotoxic Reactive Astrocytes in Human Induced Pluripotent Stem Cell Models Frontiers in Molecular Neuroscience 2022-05-03 [PMID: 35592112] (Immunohistochemistry, Human)
• YM Chung, WB Tsai, PP Khan, J Ma, JS Berek, JW Larrick, MC Hu FOXO3-dependent suppression of PD-L1 promotes anticancer immune responses via activation of natural killer cells American journal of cancer research, 2022-03-15;12(3):1241-1263. 2022-03-15 [PMID: 35411241] (IF, Human)
• Young Min Chung, Pragya P Khan, Hong Wang, Wen-Bin Tsai, Yanli Qiao, Bo Yu et al. Sensitizing tumors to anti-PD-1 therapy by promoting NK and CD8+ T cells via pharmacological activation of FOXO3 Journal for ImmunoTherapy of Cancer 2021-12-08 [PMID: 34887262] (Immunocytochemistry, Mouse)
• A Eljaafari, J Pestel, B Le Maguere, S Chanon, J Watson, M Robert, E Disse, H Vidal Adipose-Tissue-Derived Mesenchymal Stem Cells Mediate PD-L1 Overexpression in the White Adipose Tissue of Obese Individuals, Resulting in T Cell Dysfunction Cells, 2021-10-03;10(10):. 2021-10-03 [PMID: 34685625] (IHC, Human)
• A Makowska, N Lelabi, C Nothbaum, L Shen, P Busson, TTB Tran, M Eble, U Kontny Radiotherapy Combined with PD-1 Inhibition Increases NK Cell Cytotoxicity towards Nasopharyngeal Carcinoma Cells Cells, 2021-09-17;10(9):. 2021-09-17 [PMID: 34572108] (Flow Cytometry, Human)
• James W Opzoomer, Jessica A Timms, Kevin Blighe, Thanos P Mourikis, Nicolas Chapuis, Richard Bekoe et al. ImmunoCluster provides a computational framework for the nonspecialist to profile high-dimensional cytometry data eLife 2021-04-30 [PMID: 33929322] (Flow Cytometry, Human)
• T Mocan, M Ilies, I Nenu, R Craciun, A Horhat, R Susa, I Minciuna, I Rusu, LP Mocan, A Seicean, CA Iuga, NA Hajjar, M Sparchez, DC Leucuta, Z Sparchez Serum levels of soluble programmed death-ligand 1 (sPD-L1): A possible biomarker in predicting post-treatment outcomes in patients with early hepatocellular carcinoma International immunopharmacology, 2021-02-18;94(0):107467. 2021-02-18 [PMID: 33611059] (IHC, Human)
• E Acheampong, A Abed, M Morici, S Bowyer, B Amanuel, W Lin, M Millward, ES Gray Tumour PD-L1 Expression in Small-Cell Lung Cancer: A Systematic Review and Meta-Analysis Cells, 2020-10-31;9(11):. 2020-10-31 [PMID: 33142852] (IHC, Human)
• Xin Cui, Chao Ma, Varshini Vasudevaraja, Jonathan Serrano, Jie Tong, Yansong Peng et al. Dissecting the immunosuppressive tumor microenvironments in Glioblastoma-on-a-Chip for optimized PD-1 immunotherapy eLife 2020-09-10 [PMID: 32909947] (Immunocytochemistry, Human)
• Y Tian, X Zhai, A Han, H Zhu, J Yu Potential immune escape mechanisms underlying the distinct clinical outcome of immune checkpoint blockades in small cell lung cancer J Hematol Oncol, 2019-06-28;12(1):67. 6/28/2019 [PMID: 31253167]
• R Giampieri, F Piva, G Occhipinti, A Bittoni, A Righetti, S Pagliarett, A Murrone, F Bianchi, C Amantini, M Giulietti, G Ricci, G Principato, G Santoni, R Berardi, S Cascinu Clinical impact of different exosomes' protein expression in pancreatic ductal carcinoma patients treated with standard first line palliative chemotherapy PLoS ONE, 2019-05-02;14(5):e0215990. 2019-05-02 [PMID: 31048929] (ELISA Detection, Human)
• CM Cristiani, A Turdo, V Ventura, T Apuzzo, M Capone, G Madonna, D Mallardo, C Garofalo, ED Giovannone, AM Grimaldi, R Tallerico, E Marcenaro, S Pesce, G Del Zotto, V Agosti, FS Costanzo, E Gulletta, A Rizzo, A Moretta, K Kärre, PA Ascierto, M Todaro, E Carbone Accumulation of circulating CCR7+ natural killer cells marks melanoma evolution and reveals a CCL19-dependent metastatic pathway Cancer Immunol Res, 2019-04-02;0(0):. 2019-04-02 [PMID: 30940644] (ICC, IHC-P, Human)
• Mathilde Dusséaux, Guillemette Masse-Ranson, Sylvie Darche, James Ahodantin, Yan Li, Oriane Fiquet et al. Viral Load Affects the Immune Response to HBV in Mice With Humanized Immune System and Liver Gastroenterology 2017-12-01 [PMID: 28851562]
• SF Kammerer-J, A Brunot, K Bensalah, B Campillo-G, M Lefort, S Bayat, A Ravaud, F Dupuis, M Yacoub, G Verhoest, B Peyronnet, R Mathieu, A Lespagnol, J Mosser, J Edeline, B Laguerre, JC Bernhard, N Rioux-Lecl Hilar fat infiltration: A new prognostic factor in metastatic clear cell renal cell carcinoma with first-line sunitinib treatment Urol. Oncol., 2017-06-12;0(0):. 2017-06-12 [PMID: 28619630] (IHC-P, Human)
• J Zhou, KM Mahoney, A Giobbie-Hu, F Zhao, S Lee, X Liao, S Rodig, J Li, X Wu, LH Butterfiel, M Piesche, MP Manos, LM Eastman, G Dranoff, GJ Freeman, FS Hodi Soluble PD-L1 as a biomarker in malignant melanoma and checkpoint blockade Cancer Immunol Res, 2017-05-18;0(0):. 2017-05-18 [PMID: 28522460] (ELISA Development (Capture), Human)
• Stéphane Chevrier, Jacob Harrison Levine, Vito Riccardo Tomaso Zanotelli, Karina Silina, Daniel Schulz, Marina Bacac et al. An Immune Atlas of Clear Cell Renal Cell Carcinoma Cell 2017-05-04 [PMID: 28475899]
• Marlon C. Rebelatto, Anita Midha, Amita Mistry, Constantine Sabalos, Nicole Schechter, Xia Li et al. Development of a programmed cell death ligand-1 immunohistochemical assay validated for analysis of non-small cell lung cancer and head and neck squamous cell carcinoma Diagnostic Pathology 2016-12-01 [PMID: 27717372]
• Kammerer-Jacquet S, Crouzet L, Brunot A, Dagher J, Pladys A, Edeline J, Laguerre B, Peyronnet B, Mathieu R, Verhoest G, Patard J, Lespagnol A, Mosser J, Denis M, Messai Y, Gad-Lapiteau S, Chouaib S, Belaud-Rotureau M, Bensalah K, Rioux-Leclercq N Independent association of PD-L1 expression with noninactivated VHL clear cell renal cell carcinoma-A finding with therapeutic potential. Int J Cancer, 2016-09-23;140(1):142-148. 2016-09-23 [PMID: 27623354] (IHC-P, Human)
• Zoran Gatalica, Nurija Bilalovic, Juan P. Palazzo, Ryan P. Bender, Jeffrey Swensen, Sherri Z. Millis et al. Disseminated histiocytoses biomarkers beyond BRAFV600E: frequent expression of PD-L1 Oncotarget 2015-06-08 [PMID: 26110571]
• Mazel M, Jacot W, Pantel K, Bartkowiak K, Topart D, Cayrefourcq L, Rossille D, Maudelonde T, Fest T, Alix-Panabieres C Frequent expression of PD-L1 on circulating breast cancer cells. Mol Oncol, 2015-06-09;9(9):1773-82. 2015-06-09 [PMID: 26093818] (Western Blot, Human)
• Song , Xiao, Liu , Junwei, Lu , Yi, Jin , Hongchua, Huang , Dongshen Overexpression of B7-H1 correlates with malignant cell proliferation in pancreatic cancer. Oncol Rep, 2013-12-31;31(3):1191-8. 2013-12-31 [PMID: 24378899] (IHC, Human)
• Rojas D, Krishnan R IFN-gamma generates maturation-arrested dendritic cells that induce T cell hyporesponsiveness independent of Foxp3+ T-regulatory cell generation. Immunol. Lett., 2010-05-24;132(1):31-7. 2010-05-24 [PMID: 20580745] (Flow Cytometry, Human)

Bioinformatics

• Gene Symbol: CD274
• Uniprot:
  • Q9NZQ7()