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

Western blot shows lysates of NS0 mouse myeloma cell line either mock transfected or transfected with recombinant mouse 4-1BB/Fc chimera. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Mouse 4-1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF937) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF017). Specific bands were detected for 4-1BB/TNFRSF9/CD137 at approximately 50-60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Resting mouse splenocytes were stained with Rat Anti-Mouse ICOS APC-conjugated Monoclonal Antibody (Catalog # FAB168A) and either (A) Goat Anti-Mouse 4-1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF937) or (B) Normal Goat IgG Control (AB-108-C) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (F0107). View our protocol for Staining Membrane-associated Proteins.

Activated mouse splenocytes were stained with Rat Anti-Mouse ICOS APC-conjugated Monoclonal Antibody (Catalog # FAB168A) and either (A) Goat Anti-Mouse 4-1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF937) or (B) Normal Goat IgG Control (AB-108-C) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (F0107). View our protocol for Staining Membrane-associated Proteins.

4-1BB/TNFRSF9/CD137 was detected in perfusion fixed frozen sections of mouse spleen using Goat Anti-Mouse 4-1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF937) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; CTS009) and counterstained with hematoxylin (blue). Specific staining was localized to cell surfaces. View our protocol for Chromogenic IHC Staining of Frozen Tissue Sections.

NS0 cell line transfected with mouse 4‑1BB/TNFRSF9/CD137 was stained with Goat Anti-Mouse 4-1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF937, filled histogram) or Normal Goat IgG Control (AB-108-C, open histogram) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (F0107). View our protocol for Staining Membrane-associated Proteins.

Mouse 4 1BB/TNFRSF9/CD137 Antibody (Catalog # AF937) co-stimulates IFN-gamma secretion by mouse splenic T cells in the presence of anti-CD3 epsilon (MAB484). The ED50 for this effect is typically 0.400–7.00 μg/mL.

• Reactivity: Mu
• Applications: WB, Flow, IHC, CyTOF-ready, Func
• Clonality: Polyclonal
• Host: Goat
• Conjugate: Unconjugated
• Concentration: LYOPH

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

• Immunogen: Mouse myeloma cell line NS0-derived recombinant mouse 4‑1BB/TNFRSF9/CD137
  Val24-Leu187
  Accession # P20334
• Specificity: Detects mouse 4‑1BB/TNFRSF9/CD137 in direct ELISAs and Western blots. In direct ELISAs, approximately 100% cross-reactivity with recombinant rat 4-1BB/TNFRSF9 is observed. In Western blots, less than 5% cross-reactivity with recombinant human 4-1BB and recombinant mouse EDAR is observed.
• 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
  • Flow Cytometry 0.25 ug/10^6 cells
  • Immunohistochemistry 1-15 ug/mL
  • Western Blot 0.5 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 0.2 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, also known as CD137 and ILA (induced by lymphocyte activation), is a TNF receptor superfamily member and has been designated TNFRSF9. Mouse 4‑1BB cDNA encodes a 256 amino acid (aa) residues type I transmembrane protein with a putative 23 aa signal peptide, a 164 aa extracellular domain, a 21 aa transmembrane domain and a 48 aa cytoplasmic region (1‑3). A soluble 4‑1BB is released from surfaces of cells expressing the transmembrane protein (4).
Mouse 4‑1BB shares approximately 60% aa sequence identity with its human counterpart. 4‑1BB is expressed on activated CD4⁺ and CD8⁺ T cells, thymocytes, and NK cells. It is also expressed on monocytes, neutrophils, DCs and eosinophils (5). The ligand for 4‑1BB (4‑1BBL), also named TNFSF9, belongs to the TNF ligand superfamily. 4‑1BBL is predominantly expressed on activated antigen presenting cells (APCs) such as B cells, macrophages and dendritic cells (DCs). It is also expressed on most T and B lymphoma cell lines. In response to 4‑1BBL binding, 4‑1BB transduce a T cell costimulatory signal in both CD4⁺ and CD8⁺ T cells to promote survival and enhance proliferation, cytokine production and effector function. In vivo, the costimulatory activity of 4‑1BB has been shown to be important in graft-vs-host disease and antiviral CTL responses. On dendritic cells, 4‑1BB is a DC-activating molecules that enhances cytokine production and up‑regulates expression of B7-1 and B7-2 costimulatory molecules, resulting in an improved ability to stimulate T cell responses (1‑5).

1. Goodwin, R.G. et al. (1993) Eur. J. Immunol. 23:2631.
2. Alderson, M.R. et al. (1994) Eur. J. Immunol. 24:2219.
3. Kwon, B.S. and S.M. Weissman (1989) Proc. Nat. Acad. Sci. USA 86:1963.
4. Wilcox, R.A. et al. (2002) J. Immunol. 168:4262.
5. Kwon, B., H.W. Lee and B.S. Kwon, 2002, TRENDS in Immunology 23:378.

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 (AF937)(3)

Publications using AF937

• Rolando A. Garcia, James N. Roemmich, Kate J. Claycombe Evaluation of markers of beige adipocytes in white adipose tissue of the mouse Nutrition & Metabolism 12/1/2016 [PMID: 26997968]
• Miller C, Yang J, England E, Yin A, Baile C, Rayalam S Isoproterenol Increases Uncoupling, Glycolysis, and Markers of Beiging in Mature 3T3-L1 Adipocytes. PLoS ONE, 2015-09-21;10(9):e0138344. 2015-09-21 [PMID: 26390217] (IHC-Fr, Mouse)
• Palazon A, Teijeira A, Martinez-Forero I, Hervas-Stubbs S, Roncal C, Penuelas I, Dubrot J, Morales-Kastresana A, Perez-Gracia JL, Ochoa MC, Ochoa-Callejero L, Martinez A, Luque A, Dinchuk J, Rouzaut A, Jure-Kunkel M, Melero I Agonist anti-CD137 mAb act on tumor endothelial cells to enhance recruitment of activated T lymphocytes. Cancer Res., 2011-01-25;71(3):801-11. 2011-01-25 [PMID: 21266358] (IHC-Fr, Mouse)

Review for 4-1BB/TNFRSF9/CD137 Antibody (AF937) (1)

Average Rating: 4

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Verified Customer IHC-P Mouse 01/12/2015

Summary

• Application: Immunohistochemistry-Paraffin
• Sample Tested: See PMID 22799524
• Species: Mouse

Bioinformatics

• Gene Symbol: TNFRSF9
• Uniprot:
  • P20334()