4-1BB/TNFRSF9/CD137 Antibody [Unconjugated]

Immunoblot Buffer Group 1." > Immunoblot Buffer Group 1." title="Western blot shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with human 4-1BB/TNFRSF9/ CD137-eGFP Fusion. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human 4-1BB/ TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody ([catalogNumber:HAF017). A specific band was detected for 4-1BB/ TNFRSF9/CD137-eGFP Fusion at approximately 50-65 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." />

Western blot shows lysates of HEK293T human embryonic kidney cell line either mock transfected or transfected with human 4-1BB/TNFRSF9/ CD137-eGFP Fusion. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human 4-1BB/ TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody ([catalogNumber:HAF017</a>). A specific band was detected for 4-1BB/ TNFRSF9/CD137-eGFP Fusion at approximately 50-65 kDa (as indicated). This experiment was conducted under reducing conditions and using <a class=

CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." > CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." title="4-1BB/TNFRSF9/CD137 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human 4-1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to lymphocytes. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." />

4-1BB/TNFRSF9/CD137 was detected in immersion fixed paraffin-embedded sections of human tonsil using Goat Anti-Human 4-1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; <a class=

F0108) and PE-conjugated anti-human CD3 (Catalog # FAB100P). Quad marker was set based on control antibody (AB-108-C) staining." > F0108) and PE-conjugated anti-human CD3 (Catalog # FAB100P). Quad marker was set based on control antibody (AB-108-C) staining." title="Human T cells were treated for 48 hours with 5 µg/mL PHA then stained with Goat Anti-Human 4-1BB/TNFRSF9/ CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (F0108) and PE-conjugated anti-human CD3 (Catalog # FAB100P). Quad marker was set based on control antibody (AB-108-C) staining." />

Human T cells were treated for 48 hours with 5 µg/mL PHA then stained with Goat Anti-Human 4-1BB/TNFRSF9/ CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) followed by Allophycocyanin-conjugated Anti-Goat IgG Secondary Antibody (<a class=

Simple Western lane view shows lysates of HDLM‑2 human Hodgkin’s lymphoma cells, loaded at 0.2 mg/mL. A specific band was detected for 4‑1BB/TNFRSF9/CD137 at approximately 58 kDa (as indicated) using 20 µg/mL of Goat Anti-Human 4‑1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) . This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm of PHA stimulated cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." > NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm of PHA stimulated cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." title="4‑1BB/TNFRSF9/CD137 was detected in immersion fixed human untreated PBMC cells and PHA treated human PBMC cells using Goat Anti-Human 4‑1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) at 15 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm of PHA stimulated cells. View our protocol for Fluorescent ICC Staining of Cells on Coverslips." />

4‑1BB/TNFRSF9/CD137 was detected in immersion fixed human untreated PBMC cells and PHA treated human PBMC cells using Goat Anti-Human 4‑1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) at 15 µg/ml for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # <a class=

Effect of FTA on the expression of OX40, 4-1BB, and 4-1BBL in bovine PBMCs infected with BVDV.(A) Relative mRNA expression of OX40, 4-1BB, and 4-1BBL cultured with medium alone, FTA, BVDV, and BVDV plus FTA at 24, 48, 72 h in bovine PBMCs. (B) The 4-1BB protein express by western blot (left panels) and ratio of 4-1BB band intensity to that of GAPDH (right panels). 4-1BB protein in PBMCs was collected from the indicated PBMCs cultures at 24, 48, and 72 h after stimulation. Expression of GAPDH was measured as an internal control. Data are presented as the means ± SEM of three independent experiments. *P < 0.05, **P < 0.01, ***P < 0.001. Image collected and cropped by CiteAb from the following open publication (//pubmed.ncbi.nlm.nih.gov/27617959), licensed under a CC-BY license. Not internally tested by R&D Systems.

• Reactivity: Hu
• Applications: WB, Simple Western, Flow, IHC, CyTOF-ready, ELISA(Cap), Func, ICC/IF
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

4-1BB/TNFRSF9/CD137 Antibody [Unconjugated] Summary

• Immunogen: Mouse myeloma cell line NS0-derived recombinant human 4‑1BB/TNFRSF9/CD137
  Leu24-Gln186
  Accession # Q07011
• Specificity: Detects 4‑1BB/TNFRSF9/CD137 in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Goat
• Gene: TNFRSF9
• Purity Statement: Antigen Affinity-purified
• Endotoxin Note: <0.10 EU per 1 μg of the antibody by the LAL method.

Applications/Dilutions

• Dilutions:
  • Agonist Activity
  • 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)
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunocytochemistry 5-15 ug/mL
  • Immunohistochemistry 5-25 ug/mL
  • Simple Western 20 ug/mL
  • Western Blot 0.5 ug/mL
• Application Notes: ELISA Capture: Human 4-1BB/TNFRSF9/CD137 Antibody (Catalog # AF838)
  ELISA Detection: Human 4-1BB/TNFRSF9/CD137 Biotinylated Antibody (Catalog # BAF838)
  Standard: Recombinant Human 4-1BB/TNFRSF9/CD137 Fc Chimera (Catalog # 838-4B)

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 0.2 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 4-1BB/TNFRSF9/CD137 Antibody [Unconjugated]

• 4-1BB ligand receptor
• 41BB
• 4-1BB
• CD137 antigen
• CD137
• CD137MGC2172
• CDw137
• FLJ43501
• ILA
• ILAhomolog of mouse 4-1BB
• induced by lymphocyte activation (ILA)
• interleukin-activated receptor, homolog of mouse Ly63
• receptor protein 4-1BB
• T cell antigen ILA
• T-cell antigen 4-1BB homolog
• T-cell antigen ILA
• TNFRSF9
• tumor necrosis factor receptor superfamily member 9
• tumor necrosis factor receptor superfamily, member 9

Background

4-1BB is an inducible T cell surface protein belonging to the TNF receptor superfamily. It is alternatively known as TNFRSF9, CD137, and ILA. The 255 amino acid human 4-1BB is a type I transmembrane protein having in its extracellular domain four of the cysteine-rich motifs that are characteristic of the TNF receptor superfamily. The 30 kDa glycoprotein exists both as a monomer and as a dimer on T cells. The human and mouse proteins share 60% amino acid identity. 4-1BB is absent from naive T cells, but it is upregulated and continually expressed following T cell activation. The natural ligand, 4-1BBL, is a member of the TNF superfamily and is expressed on activated antigen presenting cells including dendritic cells, macrophages, and B cells. Cross-linking of 4-1BB by 4-1BBL or by agonistic antibodies transmits a potent co-stimulatory signal that enhances the effect of other activating signals such as PHA or anti-CD3 antibodies. 4-1BB signals through the
TFAF2‑NIK pathway resulting in activation of NF-kappa B and ultimately promoting proliferation and survival of T cells.

1. Vinay, D. and B. Kwon (1998) Semin. Immunol. 10:481.
2. Sica, G. and L. Chen (2000) Adv. Exp. Med. Biol. 465:355.

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 4-1BB/TNFRSF9/CD137 Antibody (AF838)(6)

Publications using AF838

• Woroniecka KI, Rhodin KE, Dechant CA et al. 4-1BB Agonism Averts TIL Exhaustion and Licenses PD-1 Blockade in Glioblastoma and Other Intracranial Cancers Clin. Cancer Res. 12/23/2019 [PMID: 31871298]
• T Masu, M Atsukawa, K Nakatsuka, M Shimizu, D Miura, T Arai, H Harimoto, C Kondo, K Kaneko, S Futagami, C Kawamoto, H Takahashi, K Iwakiri Anti-CD137 monoclonal antibody enhances trastuzumab-induced, natural killer cell-mediated cytotoxicity against pancreatic cancer cell lines with low human epidermal growth factor-like receptor 2 expression PLoS ONE, 2018-12-31;13(12):e0200664. 2018-12-31 [PMID: 30596643] (Flow Cytometry, Human)
• Hodge G, Holmes M, Jersmann H, Reynolds P, Hodge S Targeting peripheral blood pro-inflammatory cytotoxic lymphocytes by inhibiting CD137 expression: novel potential treatment for COPD. BMC Pulm Med, 2014-05-15;14(0):85. 2014-05-15 [PMID: 24885856] (Functional Assay, Human)
• Jessica Ann Hernandez-Chacon, Yufeng Li, Richard C. Wu, Chantale Bernatchez, Yijun Wang, Jeffrey S. Weber et al. Co-stimulation through the CD137/4-1BB pathway protects human melanoma tumor-infiltrating lymphocytes from activation-induced cell death and enhances anti-tumor effector function Journal of Immunotherapy 4/1/2011 [PMID: 21389874]
• Zhang H, Snyder KM, Suhoski MM, Maus MV, Kapoor V, June CH, Mackall CL 4-1BB is superior to CD28 costimulation for generating CD8+ cytotoxic lymphocytes for adoptive immunotherapy. J. Immunol., 2007-10-01;179(7):4910-8. 2007-10-01 [PMID: 17878391] (Functional Assay, Human)
• Maerten P, Geboes K, De Hertogh G, Shen C, Cadot P, Bullens DM, Van Assche G, Penninckx F, Rutgeerts P, Ceuppens JL Functional expression of 4-1BB (CD137) in the inflammatory tissue in Crohn&#039;s disease. Clin. Immunol., 2004-09-01;112(3):239-46. 2004-09-01 [PMID: 15308117] (Functional Assay, IHC-Fr, Human)

Bioinformatics

• Gene Symbol: TNFRSF9
• Uniprot:
  • Q07011()