Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The purified MABTNFR1-A1 antibody (Cat. No. 552535) is useful as a capture antibody for a sandwich ELISA which measures TNFRI protein levels. Purified MABTNFR1-A1 antibody can be paired with the biotinylated MABTNFR1-B1 anti-human TNFRI (Cat. No. 550900) as the detecting antibody, with recombinant soluble human TNFRI as the standard. Purified MABTNFR1-A1 antibody should be titrated between 1 - 4 µg/ml to determine the optimal coating concentration for ELISA. To obtain linear standard curves, doubling dilutions of recombinant human TNFRI ranging from 2000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, visit the online protocols, or the chapter on ELISA in the Immune Function Handbook, both of which are on our web site at www.bdbiosciences.com..
Note: This ELISA antibody pair shows no cross-reactivity with the following recombinant human cytokines: IL-1RII, IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p40, IL-12p70, IL-13, IL-15, TNF, Lt-α, TNFRII, GM-CSF, TGFβ, This ELISA antibody pair also shows no cross-reactivity with recombinant cytokines from other species: rat TNF, mouse TNF, mouse TNFRI, mouse TNFRII.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
The MABTNFR1-A1 antibody reacts with the extracellular domain of the 55 kDa receptor for the human cytokines: TNF (aka tumor necrosis factor-alpha , TNF-α) and lymphotoxin-alpha (LT-α3, aka, lymphotoxin or TNF-β). This receptor is referred to as the p55 or Type I Tumor Necrosis Factor Receptor (TNFRI) or CD120a. TNFRI are expressed by a variety of cell lines, tumor cells, and normal cell types including T cells, monocytes, macrophages, neutrophils, endothelial cells, hepatocytes, chondrocytes, and fibroblasts. Naive B cells express very low or undetectable levels of TNFRI whereas mature erythrocytes and platelets are uniformly negative for TNFRI expression. For the generation of the MABTNFRI-B1 hybridoma, BALB/c mice were immunized with purified, full-length human TNFRI protein. MABTNFRI-A1 specifically binds to natural and recombinant truncated forms of TNFRI.
Development References (9)
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Aggarwal S, Gollapudi S, Gupta S. Increased TNF-alpha-induced apoptosis in lymphocytes from aged humans: changes in TNF-alpha receptor expression and activation of caspases. J Immunol. 1999; 162(4):2154-2161. (Biology). View Reference
-
Brockhaus M, Schoenfeld HJ, Schlaeger EJ, Hunziker W, Lesslauer W, Loetscher H. Identification of two types of tumor necrosis factor receptors on human cell lines by monoclonal antibodies. Proc Natl Acad Sci U S A. 1990; 87(8):3127-3131. (Biology). View Reference
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Browning JL, Dougas I, Ngam-ek A, et al. Characterization of surface lymphotoxin forms. Use of specific monoclonal antibodies and soluble receptors.. J Immunol. 1995; 154(1):33-46. (Biology). View Reference
-
Erikstein BK, Smeland EB, Blomhoff HK, et al. Independent regulation of 55-kDa and 75-kDa tumor necrosis factor receptors during activation of human peripheral blood B lymphocytes. Eur J Immunol. 1991; 21(4):1033-1037. (Biology). View Reference
-
Gehr G, Gentz R, Brockhaus M, Loetscher H, Lesslauer W. Both tumor necrosis factor receptor types mediate proliferative signals in human mononuclear cell activation. J Immunol. 1992; 149(3):911-917. (Biology). View Reference
-
Heilig B, Mapara M, Brockhaus M, Krauth K, Dörken B. Two types of TNF receptors are expressed on human normal and malignant B lymphocytes. Clin Immunol Immunopathol. 1991; 61(2):260-267. (Biology). View Reference
-
Hohmann HP, Remy R, Brockhaus M, van Loon AP. Two different cell types have different major receptors for human tumor necrosis factor (TNF alpha). J Biol Chem. 1989; 264(25):14927-14934. (Biology). View Reference
-
Wallach D, Engelmann H, Nophar Y, et al. Soluble and cell surface receptors for tumor necrosis factor. Agents Actions Suppl. 1991; 35:51-57. (Biology). View Reference
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Zola H. Detection of cytokine receptors by flow cytometry. In: Coligan JE, Kruisbeek AM, Margulies DH, Shevach EM, Strober W, ed. Current Protocols in Immunology. New York: Green Publishing Associates and Wiley-Interscience; 1995:6.21.1-6.21.18.
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
