Human DcR3/TNFRSF6B Antibody

AF142 1 Citation Datasheet / COA / SDS

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AF142
AF142-SP

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Human DcR3/TNFRSF6B Antibody in Immunohistochemistry

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Human DcR3/TNFRSF6B Antibody Summary

Species Reactivity

Human

Specificity

Detects human DcR3/TNFRSF6B in ELISAs and Western blots. In direct ELISAs, less than 1% cross-reactivity with recombinant human (rh) TRAIL R1, rhTRAIL R2, rhTRAIL R3, and rhTRAIL R4 is observed.

Source

Polyclonal Goat IgG

Purification

Antigen Affinity-purified

Immunogen

S. frugiperda insect ovarian cell line Sf 21-derived recombinant human DcR3/TNFRSF6B
Val24-His300
Accession # O95407

Formulation

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

Endotoxin Level

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

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

0.1 µg/mL

Recombinant Human DcR3/TNFRSF6B Fc Chimera (Catalog # 142-DC)

Immunohistochemistry

5-15 µg/mL

See below

Neutralization

Measured by its ability to neutralize DcR3/TNFRSF6B-mediated inhibition of apoptosis in the Jurkat human acute T cell leukemia cell line. The Neutralization Dose (ND₅₀) is typically 0.1-0.5 µg/mL in the presence of 50 ng/mL Recombinant Human DcR3/TNFRSF6B Fc Chimera, 2 ng/mL Recombinant Human Fas Ligand/TNFSF6.

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Data Examples

Immunohistochemistry

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DcR3/TNFRSF6B in Human Stomach Cancer Tissue. DcR3/TNFRSF6B was detected in immersion fixed paraffin-embedded sections of human stomach cancer using 10 µg/mL Goat Anti-Human DcR3/TNFRSF6B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF142) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the membrane of epithelial cells in gastric glands. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Neutralization

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DcR3/TNFRSF6B Inhibition of Fas Ligand/TNFSF6-induced Apoptosis and Neutralization by Human DcR3/TNFRSF6B Antibody. In the presence of a cross-linking antibody, Mouse polyHistidine Monoclonal Antibody (10 µg/mL, Catalog # MAB050), Recombinant Human DcR3/TNFRSF6B Fc Chimera (Catalog # 142-DC) inhibits Recombinant Human Fas Ligand/TNFSF6 (Catalog # 126-FL) induced apoptosis in the Jurkat human acute T cell leukemia cell line in a dose-dependent manner (orange line). Under these conditions, inhibition of Recombinant Human Fas Ligand/TNFSF6 (2 ng/mL) activity elicited by Recombinant Human DcR3/TNFRSF6B Fc Chimera (50 ng/mL) is neutralized (green line) by increasing concentrations of Goat Anti-Human DcR3/TNFRSF6B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF142). The ND₅₀ is typically 0.1-0.5 µg/mL.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS.

Reconstitution Buffer Available

Reconstitution Buffer 1 (PBS)

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

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: DcR3/TNFRSF6B

Human decoy receptor 3 (DcR3), also called TNFRSF6B, TR6, and M68, is a member of the TNF receptor superfamily. The cDNA of DcR3 encodes a 300 amino acid (aa) polypeptide with a putative 23 aa signal peptide. Like osteoprotegerin (OPG), DcR3 lacks a transmembrane sequence and is a secreted protein. DcR3 shares sequence identity with OPG (31%), TNF RII (29%), and Fas (17%). It was found to be expressed in a variety of different tissues and at high levels in many malignant tumors. Among TNF family members, DcR3 was shown to bind with Fas ligand (FasL) and LIGHT and inhibit FasL- and LIGHT-induced apoptosis. Thus, in addition to DcR1, DcR2, and OPG, DcR3 is another TNFR family molecule which modulates ligands that induce apoptosis. Over-expression of DcR3 might be a mechanism by which certain tumors escape immune-cytotoxic attack.

References

Pitti, R.M. et al. (1998) Nature 396:699.
Yu, K-Y. et al. (1999) J. Biochem. Chem. 274:13733.
Bai, C. et al. (2000) Proc. Natl. Acad. Sci. USA 97:1230.

Long Name

Decoy Receptor 3

Entrez Gene IDs

8771 (Human); 107126607 (Cynomolgus Monkey)

Alternate Names

DcR3; DCR3Decoy receptor for Fas ligand; Decoy receptor 3; M68; M68DJ583P15.1.1; TNFRSF6B; TR6; TR6DcR3; tumor necrosis factor receptor superfamily member 6B; tumor necrosis factor receptor superfamily, member 6b, decoy

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Citation for Human DcR3/TNFRSF6B Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

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