Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Flow Cytometry, Immunocytochemistry, Simple Western, CyTOF-ready

Cited:

Immunocytochemistry

Label

Unconjugated

Antibody Source

Recombinant Monoclonal Rabbit IgG Clone # 2356B
View all 4-1BB/TNFRSF9/CD137 Primary Antibodies »

Product Specifications

Immunogen

Chinese hamster ovary cell line CHO-derived recombinant human 4‑1BB/TNFRSF9/CD137
Leu24-His183
Accession # Q07011

Specificity

Detects human 4‑1BB/TNFRSF9/CD137 in direct ELISAs.

Clonality

Monoclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Human 4‑1BB/TNFRSF9/CD137 Antibody

Detection of Human 4-1BB/TNFRSF9/CD137 antibody by Western Blot.

Detection of Human 4‑1BB/TNFRSF9/CD137 by Western Blot.

Western blot shows lysates of HDLM-2 human Hodgkin's lymphoma cell line and human tonsil tissue. PVDF membrane was probed with 2 µg/mL of Rabbit Anti-Human 4-1BB/TNFRSF9/CD137 Monoclonal Antibody (Catalog # MAB8381) followed by HRP-conjugated Anti-Rabbit IgG Secondary Antibody (HAF008). Specific bands were detected for 4-1BB/TNFRSF9/CD137 at approximately 32 and 40 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Detection of 4-1BB/TNFRSF9/CD137 antibody in HEK293 Human Cell Line Transfected with Human 4-1BB/TNFRSF9/CD137 and eGFP antibody by Flow Cytometry.

Detection of 4-1BB/TNFRSF9/CD137 in HEK293 Human Cell Line Transfected with Human 4-1BB/TNFRSF9/CD137 and eGFP by Flow Cytometry.

HEK293 human embryonic kidney cell line transfected with either (A) human 4-1BB/TNFRSF9/CD137 or (B) irrelevant protein, and eGFP was stained with Rabbit Anti-Human 4-1BB/TNFRSF9/CD137 Monoclonal Antibody (Catalog # MAB8381) followed by APC-conjugated Goat-anti Rabbit IgG secondary antibody (F0111). Quadrant markers were set based on control antibody staining (MAB1050). View our protocol for Staining Membrane-associated Proteins.

4-1BB/TNFRSF9/CD137 antibody in Human PBMCs and HEK293 Cell Line by Immunocytochemistry (ICC).

4‑1BB/TNFRSF9/CD137 in Human PBMCs and HEK293 Cell Line.

4-1BB/TNFRSF9/CD137 was detected in immersion fixed peripheral blood mononuclear cells (PBMCs) treated with PHA (positive control, left panel) and HEK293 human embryonic kidney cell line (negative control, right panel) using Rabbit Anti-Human 4-1BB/TNFRSF9/CD137 Monoclonal Antibody (Catalog # MAB8381) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Rabbit IgG Secondary Antibody (red NL004) and counterstained with DAPI (blue). Specific staining was localized to cell surfaces. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Detection of Human 4‑1BB/TNFRSF9/CD137 by Simple WesternTM.

Simple Western lane view shows lysates of HDLM‑2 human Hodgkin’s lymphoma cells, loaded at 0.2 mg/mL. Specific bands were detected for 4‑1BB/TNFRSF9/CD137 at approximately 49 and 57 kDa (as indicated) using 20 µg/mL of Rabbit Anti-Human 4‑1BB/TNFRSF9/CD137 Monoclonal Antibody (Catalog # MAB8381). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Applications for Human 4‑1BB/TNFRSF9/CD137 Antibody

Application
Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

0.25 µg/106 cells
Sample: HEK293 Human Cell Line Transfected with Human 4-1BB/TNFRSF9/CD137 and eGFP

Immunocytochemistry

8-25 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells (PBMCs) and HEK293 human embryonic kidney cell line

Simple Western

20 µg/mL
Sample: HDLM‑2 human Hodgkin's lymphoma cells

Western Blot

2 µg/mL
Sample: HDLM‑2 human Hodgkin's lymphoma cell line and human tonsil tissue

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Advanced Features

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Protein A or G purified from cell culture supernatant

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & 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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: 4-1BB/TNFRSF9/CD137

4-1BB, also known as CD137 and TNFRSF9, is an approximately 30 kDa transmembrane glycoprotein in the TNF receptor superfamily. 4-1BB functions in the development and activation of multiple immune cells (1). Mature human 4-1BB consists of a 163 amino acid (aa) extracellular domain (ECD) with four TNFR cysteine-rich repeats, a 27 aa transmembrane segment, and a 42 aa cytoplasmic domain (2, 3). Within the ECD, human 4-1BB shares 60% aa sequence identity with mouse and rat 4-1BB. 4-1BB is expressed as a disulfide-linked homodimer on various populations of activated T cell including CD4+, CD8+, memory CD8+, NKT, and regulatory T cells (4-7) as well as on myeloid and mast cell progenitors, dendritic cells, mast cells, and bacterially infected osteoblasts (8-11). It binds with high affinity to the transmembrane 4-1BB Ligand/TNFSF9 which is expressed on antigen presenting cells and myeloid progenitor cells (3, 8). This interaction costimulates the proliferation, activation, and/or survival of the 4-1BB expressing cell (3-7). It can also enhance the activation-induced cell death of repetitively stimulated T cells (3). Mice lacking 4-1BB show augmented T cell activation, perhaps due to its absence on regulatory T cells (12). 4-1BB can associate with OX40 on activated T cells, forming a complex that responds to either ligand and inhibits Treg and CD8+ T cell proliferation (13). Reverse signaling through 4-1BB Ligand inhibits the development of dendritic cells, B cells, and osteoclasts (8, 11) but supports mature dendritic cell survival and costimulates the proliferation and activation of mast cells (9, 10).
4-1BB activation enhances CD8+ T cell and NK cell mediated anti-tumor immunity (14). It also contributes to the development of inflammation in high fat diet-induced metabolic syndrome (15). Soluble forms of 4-1BB and 4-1BB Ligand circulate at elevated levels in the serum of rheumatoid arthritis and hematologic cancer patients, respectively (16, 17).

References

  1. Wang, C. et al. (2009) Immunol. Rev. 229:192.
  2. Schwarz, H. et al. (1993) Gene 134:295.
  3. Alderson, M.R. et al. (1994) Eur. J. Immunol. 24:2219.
  4. Wen, T. et al. (2002) J. Immunol. 168:4897.
  5. Pulle, G. et al. (2006) J. Immunol. 176:2739.
  6. Zheng, G. et al. (2004) J. Immunol. 173:2428.
  7. Kim, D. et al. (2008) J. Immunol. 180:2062.
  8. Lee, S. et al. (2008) Nat. Immunol. 9:917.
  9. Choi, B.K. et al. (2009) J. Immunol. 182:4107.
  10. Nishimoto, H. et al. (2005) Blood 106:4241.
  11. Saito, K. et al. (2004) J. Biol. Chem. 279:13555.
  12. Lee, S. et al. (2005) J. Immunol. 174:6803.
  13. Ma, B.Y. et al. (2005) Blood 106:2002.
  14. Choi, B.K. et al. (2010) J. Immunol. 185:1404.
  15. Kim, C. et al. (2011) Diabetes 60:3159.
  16. Michel, J. et al. (1998) Eur. J. Immunol. 28:290.
  17. Salih, H.R. et al. (2001) J. Immunol. 167:4059.

Alternate Names

41BB, CD137, ILA, TNFRSF9

Entrez Gene IDs

3604 (Human); 21942 (Mouse); 500590 (Rat); 102127961 (Cynomolgus Monkey)

Gene Symbol

TNFRSF9

UniProt

Q07011

Additional 4-1BB/TNFRSF9/CD137 Products

Product Documents for Human 4‑1BB/TNFRSF9/CD137 Antibody

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Product Specific Notices for Human 4‑1BB/TNFRSF9/CD137 Antibody

For research use only

Citations for Human 4‑1BB/TNFRSF9/CD137 Antibody

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Protocols

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