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Rat 4-1BB/TNFRSF9/CD137 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF9029

R&D Systems, part of Bio-Techne
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AF9029-100
AF9029-SP

Key Product Details

Validated by

Biological Validation

Species Reactivity

Rat

Applications

CyTOF-ready, Flow Cytometry, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG

Product Specifications

Immunogen

Mouse myeloma cell line NS0-derived recombinant rat 4-1BB.

Specificity

Detects rat 4-1BB in direct ELISAs.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Rat 4-1BB/TNFRSF9/CD137 Antibody

Detection of 4-1BB/TNFRSF9/CD137 antibody in Rat Splenocytes antibody by Flow Cytometry.

Detection of 4‑1BB/TNFRSF9/CD137 in Rat Splenocytes by Flow Cytometry.

Rat splenocytes either (A) resting or (B) activated with with 50 ng/mL PMA and 200 ng/mL Calcium Ionomycin overnight were stained with Goat Anti-Rat 4-1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF9029) followed by Phycoerythrin-conjugated Anti-Goat IgG Secondary Antibody (Catalog # F0107) and APC-conjugated Anti-Rat CD3. Quadrant markers were set based on control antibody staining (Catalog # AB-108-C). View our protocol for Staining Membrane-associated Proteins.
4-1BB/TNFRSF9/CD137 antibody in Rat Splenocytes by Immunocytochemistry (ICC).

4‑1BB/TNFRSF9/CD137 in Rat Splenocytes.

4-1BB/TNFRSF9/CD137 was detected in immersion fixed rat splenocytes treated with calcium ionomycin and PMA using Goat Anti-Rat 4-1BB/TNFRSF9/ CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF9029) 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. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Applications for Rat 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: Rat splenocytes treated with PMA and Calcium Ionomycin

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed rat splenocytes

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Reconstitution Buffer Available:
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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.

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 rat consists of a 166 amino acid (aa) extracellular domain (ECD) with four TNFR cysteine‑rich repeats, a 21 aa transmembrane segment, and a 48 aa cytoplasmic domain. Within the ECD, rat 4‑1BB shares 60% and 79% aa sequence identity with human and mouse 4‑1BB, respectively. 4‑1BB is expressed as a disulfide‑linked homodimer on various populations of activated T cells including CD4+, CD8+, memory CD8+, NKT, and regulatory T cells (2‑5) as well as on myeloid and mast cell progenitors, dendritic cells, mast cells, and bacterially infected osteoblasts (6‑9). It binds with high affinity to the transmembrane 4‑1BB Ligand/TNFSF9 which is expressed on antigen presenting cells and myeloid progenitor cells (6, 10). This interaction co‑stimulates the proliferation, activation, and/or survival of the 4‑1BB expressing cell (2‑5, 10). It can also enhance the activation‑induced cell death of repetitively stimulated T cells (10). Mice lacking 4‑1BB show augmented T cell activation, perhaps due to its absence on regulatory T cells (11). 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 (12). Reverse signaling through 4‑1BB Ligand inhibits the development of dendritic cells, B cells, and osteoclasts (6, 9) but supports mature dendritic cell survival and co‑stimulates the proliferation and activation of mast cells (7, 8). 4‑1BB activation enhances CD8+ T cell and NK cell mediated anti‑tumor immunity (13). It also contributes to the development of inflammation in high fat diet‑induced metabolic syndrome (14). Soluble forms of 4‑1BB and 4‑1BB Ligand circulate at elevated levels in the serum of rheumatoid arthritis and hematologic cancer patients, respectively (15, 16).

References

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

Additional 4-1BB/TNFRSF9/CD137 Products

Product Documents for Rat 4-1BB/TNFRSF9/CD137 Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Rat 4-1BB/TNFRSF9/CD137 Antibody

For research use only

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