Human 4-1BB/TNFRSF9/CD137 Antibody

R&D Systems | Catalog # AF838

R&D Systems

Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, ELISA Capture (Matched Antibody Pair), Flow Cytometry, Immunocytochemistry, Simple Western, Agonist Activity, CyTOF-ready

Cited:

Immunohistochemistry-Frozen, Flow Cytometry, Functional Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all 4-1BB/TNFRSF9/CD137 Primary Antibodies »

Product Specifications

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.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Endotoxin Level

<0.10 EU per 1 μg of the antibody by the LAL method.

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 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.
4-1BB/TNFRSF9/CD137 antibody in Human Tonsil by Immunohistochemistry (IHC-P).

4‑1BB/TNFRSF9/CD137 in Human Tonsil.

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.
Detection of 4-1BB/TNFRSF9/CD137 antibody in PHA-treated Human T Cells antibody by Flow Cytometry.

Detection of 4‑1BB/TNFRSF9/CD137 in PHA-treated Human T Cells by Flow Cytometry.

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.

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. 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.

Detection of 4‑1BB/TNFRSF9/CD137 in Human Untreated PBMC and PHA Treated Human PBMC.

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

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

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 (https://pubmed.ncbi.nlm.nih.gov/27617959), licensed under a CC-BY license. Not internally tested by R&D Systems.
Human 4-1BB / TNFRSF9 / CD137 ELISA Standard Curve

Human 4-1BB / TNFRSF9 / CD137 ELISA Standard Curve

Recombinant Human 4‑1BB/TNFRSF9/CD137 Fc Chimera (Catalog # 838-4B) was serially diluted and captured by Goat Anti-Human 4‑1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human 4‑1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Human 4-1BB / TNFRSF9 / CD137 ELISA Standard Curve

Human 4-1BB / TNFRSF9 / CD137 ELISA Standard Curve

Recombinant Human 4‑1BB/TNFRSF9/CD137 Fc Chimera (Catalog # 838-4B) was serially diluted and captured by Goat Anti-Human 4‑1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human 4‑1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Human 4-1BB / TNFRSF9 / CD137 ELISA Standard Curve

Human 4-1BB / TNFRSF9 / CD137 ELISA Standard Curve

Recombinant Human 4‑1BB/TNFRSF9/CD137 Fc Chimera (Catalog # 838-4B) was serially diluted and captured by Goat Anti-Human 4‑1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human 4‑1BB/TNFRSF9/CD137 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF838) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)

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

Application
Recommended Usage

Agonist Activity

Measured by its ability to co-stimulate IFN-gamma secretion by human T cells in the presence of anti-CD3.
The ED50 for this effect is typically ≤ 20 μg/mL.

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: Human T cells treated with PHA

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed human peripheral blood mononuclear cells

Immunohistochemistry

5-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human tonsil

Simple Western

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

Western Blot

0.5 µg/mL
Sample: HEK293T human embryonic kidney cell line transfected with human 4-1BB/TNFRSF9/CD137-eGFP Fusion

Human 4-1BB/TNFRSF9/CD137 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 0.2-0.8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human 4‑1BB/TNFRSF9/CD137 Biotinylated Antibody (Catalog # BAF838)
  • Standard: Recombinant Human 4-1BB/TNFRSF9 Fc Chimera Protein, CF (Catalog # 838-4B)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 1 review rated 5 using AF838 in the following applications:

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Flow Cytometry

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.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.<br/>*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 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.

References

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

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

Certificate of Analysis

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

Note: Certificate of Analysis not available for kit components.

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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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  • Human 4-1BB/TNFRSF9/CD137 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Adult kidney
    Species: Human
    Verified Customer | Posted 10/13/2017

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