Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, Neutralization

Cited:

Immunohistochemistry-Paraffin, Immunohistochemistry-Frozen, Western Blot, Neutralization, Flow Cytometry, Immunocytochemistry, Immunoprecipitation, Bioassay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all TRAILR1/TNFRSF10A Primary Antibodies »

Product Specifications

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human TRAIL R1/TNFRSF10A
Ala24-Asn239
Accession # AAC51226

Specificity

Detects human TRAIL R1/TNFRSF10A in direct ELISAs and Western blots. In Western blots, approximately 10% cross-reactivity with recombinant human (rh) TRAIL R2 is observed and less than 1% cross-reactivity with rhTRAIL R3 and rhTRAIL R4 is observed.

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 TRAILR1/TNFRSF10A Antibody

Detection of Human TRAIL R1/TNFRSF10A antibody by Western Blot.

Detection of Human TRAIL R1/TNFRSF10A by Western Blot.

Western blot shows lysates of TF-1 human erythroleukemic cell line. PVDF membrane was probed with 1 µg/mL of Goat Anti-Human TRAIL R1/TNFRSF10A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF347) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF019). A specific band was detected for TRAIL R1/TNFRSF10A at approximately 50 kDa (as indicated). This experiment was conducted under reducing conditions.

TRAIL R1/TNFRSF10A Inhibition of TRAIL/TNFSF10-induced Cytotoxicity and Neutralization by Human TRAIL R1/TNFRSF10A Antibody.

TRAIL R1/TNFRSF10A Inhibition of TRAIL/TNFSF10-induced Cytotoxicity and Neutralization by Human TRAIL R1/TNFRSF10A Antibody.

In the presence of a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody (Catalog # MAB050) and the metabolic inhibitor actinomycin D (1 µg/mL), Recombinant Human TRAIL R1/TNFRSF10A Fc Chimera (Catalog # 347-DR) inhibits Recombinant Human TRAIL/TNFSF10 (Catalog # 375-TL) induced cytotoxicity in the L-929 mouse fibroblast cell line in a dose-dependent manner (orange line), as measured by crystal violet staining. Under these conditions, inhibition of Recombinant Human TRAIL/TNFSF10 (12 ng/mL) activity elicited by Recombinant Human TRAIL R1/TNFRSF10A Fc Chimera (10 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human TRAIL R1/ TNFRSF10A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF347). The ND50 is typically 0.02-0.055 µg/mL.

TRAIL R1/TNFRSF10A antibody in Human Brain by Immunohistochemistry (IHC-P).

TRAIL R1/TNFRSF10A in Human Brain.

TRAIL R1/TNFRSF10A was detected in immersion fixed paraffin-embedded sections of human brain using Goat Anti-Human TRAIL R1/TNFRSF10A Antigen Affinity-purified Polyclonal Antibody (Catalog # AF347) at 10 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm in neurons. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human TRAILR1/TNFRSF10A by Western Blot

Detection of Human TRAILR1/TNFRSF10A by Western Blot

B3GNT2 disrupts ligand–receptor interactions between tumor and T cells.a Cell survival against T cell cytotoxicity (top) and T cell IFN gamma secretion (bottom) in A375 cells overexpressing B3GNT2 or GFP that have been treated with different concentrations of kifunensine. Kifunensine was used to pretreat A375 cells and was present during co-culture with T cells at E:T ratio of 3. Kifunensine-treated cells that were co-cultured with ESO T cells were compared to kifunensine-treated cells cultured in parallel without T cells to determine percent survival. N = 6. Two-tailed t tests were performed. b Tomato lectin IP of A375 cells overexpressing GFP or B3GNT2 followed by western blot for different B3GNT2 target proteins. 2% of the input and no lectin IP controls are shown. Data are representative of two independent experiments. c Western blots of A375 cells overexpressing B3GNT2 or GFP that were treated with kifunensine (KIF) or benzyl-2-acetamido-2-deoxy-alpha -D-galactopyranoside (BAG) to remove N- or O-glycosylation respectively. Data are representative of two independent experiments. d Histograms (top) and corresponding median fluorescence intensity (MFI; bottom) showing binding of recombinant T cell proteins to A375 cells measured by flow cytometry. A375 cells were overexpressing GFP or B3GNT2 and treated with KIF or BAG. N = 3. Two-tailed t tests were performed. e Schematic showing the tumor cell surface ligands and receptors modified by B3GNT2 to disrupt interactions with T cells that mediate cytotoxicity. All values are mean ± s.e.m. ns not significant. Source data are provided in Source Data 4. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35338135), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human TRAILR1/TNFRSF10A Antibody

Application
Recommended Usage

Immunohistochemistry

5-15 µg/mL
Sample:  Immersion fixed paraffin-embedded sections of human brain

Western Blot

1 µg/mL
Sample: TF‑1 human erythroleukemic cell line

Neutralization

Measured by its ability to neutralize TRAIL R1/TNFRSF10A-mediated inhibition of cytotoxicity in the L‑929 mouse fibroblast cell line. The Neutralization Dose (ND50) is typically 0.02-0.055 µg/mL in the presence of 10 ng/mL Recombinant Human TRAIL R1/TNFRSF10A Fc Chimera, 12 ng/mL Recombinant Human TRAIL/TNFSF10, a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody, and 1 µg/mL actinomycin D.

Reviewed Applications

Read 1 review rated 4 using AF347 in the following applications:

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. *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: TRAILR1/TNFRSF10A

Human TRAIL R1, also called DR4, is a type 1, TNF R family, membrane protein which is a receptor for TRAIL (APO2 ligand). In the TNF superfamily nomenclature, TRAIL RI is referred to as TNFRSF10A. TRAIL R1 cDNA encodes a 468 amino acid residue precursor protein containing extracellular cysteine-rich domains, a transmembrane domain and a cytoplasmic death domain. Among the TNF receptor family proteins, TRAIL R1 is most closely related to TRAIL R2/DR5, sharing 55% amino acid sequence identity. Binding of trimeric TRAIL to TRAIL R1 induces apoptosis. The induction of apoptosis likely requires oligomerization of the receptor. The human TRAIL R1/Fc chimera neutralizes the ability of TRAIL to induce apoptosis. Besides TRAIL R1, an additional TRAIL R2/DR5, which tranduces apoptosis signal, and two TRAIL decoy receptors, which antagonize TRAIL-induced apoptosis, have been reported.

References

  1. Pan, G. et al. (1997) Science 276:111.
  2. Golstein, P. (1997) Curr. Biol. 7:R750.

Long Name

TRAIL Receptor 1

Alternate Names

CD261, DR4, TNFRSF10A, TRAIL R1

Entrez Gene IDs

8797 (Human)

Gene Symbol

TNFRSF10A

UniProt

AAC51226

Additional TRAILR1/TNFRSF10A Products

Product Documents for Human TRAILR1/TNFRSF10A Antibody

Certificate of Analysis

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Product Specific Notices for Human TRAILR1/TNFRSF10A Antibody

For research use only

Citations for Human TRAILR1/TNFRSF10A Antibody

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  • Human TRAIL R1/TNFRSF10A Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: HT-29 human colon adenocarcinoma cell line and MCF-7 human breast cancer cell line
    Species: Human
    Verified Customer | Posted 07/23/2018
    Total cell lysates from HT29 and MCF-7 were subjected to western blot. PVDF membrane were probed with 1 mm/ml Human DR4 Antibody (AF347). A specific band was detected for DR4 at approximately 55 kDa. This experiment was conducted under reducing conditions.
    Human TRAILR1/TNFRSF10A Antibody AF347

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