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      Biotin Mouse Anti-Human CD120a
      Biotin Mouse Anti-Human CD120a
      Flow cytometric analysis of expression of cell surface TNFRI by whole-lysed human blood. Whole human blood was stained with Biotin Mouse Anti-Human CD120a (Cat No. 550900) followed by PE Streptavidin (Cat. No. 554061). Red blood cells were subsequently lysed with Lysing Buffer (Cat No. 555899). Staining with the Biotin Mouse Anti-Human CD120a (filled histogram) is compared to staining obtained using the Biotin Mouse IgG2a, κ Isotype Control (Cat. No. 553455; open histograms). Histograms in figure are gated on the lymphocyte (left panel), monocyte (middle panel) and granulocyte (right panel) populations based on the side and forward light scattering characteristics. Note: Certain human cell lines or cell types (e.g., neutrophils, monocytes) can first be treated with reagents that block receptors for the Fc regions of immunoglobulin to avoid nonspecific immunofluorescent staining mediated by Fc receptors (for example see Reference by Browning et. al.).
      Biotin Mouse Anti-Human CD120a
      Flow cytometric analysis of expression of cell surface TNFRI by whole-lysed human blood. Whole human blood was stained with Biotin Mouse Anti-Human CD120a (Cat No. 550900) followed by PE Streptavidin (Cat. No. 554061). Red blood cells were subsequently lysed with Lysing Buffer (Cat No. 555899). Staining with the Biotin Mouse Anti-Human CD120a (filled histogram) is compared to staining obtained using the Biotin Mouse IgG2a, κ Isotype Control (Cat. No. 553455; open histograms). Histograms in figure are gated on the lymphocyte (left panel), monocyte (middle panel) and granulocyte (right panel) populations based on the side and forward light scattering characteristics. Note: Certain human cell lines or cell types (e.g., neutrophils, monocytes) can first be treated with reagents that block receptors for the Fc regions of immunoglobulin to avoid nonspecific immunofluorescent staining mediated by Fc receptors (for example see Reference by Browning et. al.).
      Product Details
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      BD Pharmingen™
      TNF Receptor Type I; TNFR1; TNFR55; TNFR60; TNFRSF1A
      Human (QC Testing)
      Mouse BALB/c IgG2a, κ
      Human CD120a Recombinant Protein
      Flow cytometry (Routinely Tested), ELISA Detection (Tested During Development)
      0.5 mg/ml
      AB_393945
      Aqueous buffered solution containing ≤0.09% sodium azide.
      RUO


      Preparation And Storage

      Store undiluted at 4°C. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed.
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      Recommended Assay Procedures

      Immunofluorescent Staining and Flow Cytometric Analysis: Biotin Mouse Anti-Human CD120a (Cat. No. 550900) can be used for immunofluorescent staining (≤ 1 µg antibody/10×6 cells) and flow cytometric analysis of nucleated human cells to measure their expressed levels of surface TNFRI. A two-layer staining protocol is recommended for maximizing the detection of TNFRI expressed by cells as detailed in the figure legend. As a demonstration of specificity, the binding of MABTNFR1-B1 to TNFRI is inhibited when human TNFRI+ target cells are preincubated with saturating levels of recombinant human TNF at 4°C, i.e., when the TNFRI are bound with ligand. Based on our data, a recombinant human TNF level above 50 ng/10×6 cells is sufficient to completely inhibit the binding of MABTNFR1-B1 (0.06 µg/10×6 cells). Please note also that as a consequence of in vivo or in vitro activation, cell surface TNFRI can either be shed by cells or transiently expressed at higher levels. As a result, cellular activation can affect the cell's overall expressed level of surface TNFRI.

      ELISA Detection: Biotin Mouse Anti-Human CD120a serves as the detection antibody in a sandwich ELISA for measuring human soluble TNFRI protein levels. Biotin Mouse Anti-Human CD120a antibody can be paired with the Purified Mouse Anti-Human CD120a (Cat No. 552535) using recombinant human soluble TNFRI as the standard. This detection antibody should be titrated between 0.5 - 1 µg/ml to determine its optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of recombinant soluble human TNFRI ranging from 200 to 15 pg/ml are recommended for inclusion in each ELISA plate.  For specific methodology, please visit the protocols section under "ELISA and ELISPOT" at our website, http://www.bdbiosciences.com/us/s/resources.

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      Product Notices

      1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
      2. An isotype control should be used at the same concentration as the antibody of interest.
      3. 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.
      4. For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
      5. Please refer to http://regdocs.bd.com to access safety data sheets (SDS).
      6. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
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      552536 Rev. 2
      Antibody Details
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      MABTNFR1-B1

      The TNFR1-B1 monoclonal antibody specifically recognizes the extracellular domain of CD120a. CD120a is encoded by TNFRSF1A (Tumor necrosis factor receptor superfamily member 1A). CD120a is also known as the 55 kDa receptor for the human cytokines, tumor necrosis factor (TNF) and lymphotoxin-alpha (LT-a). This receptor is referred to as the p55 or Type I Tumor Necrosis Factor Receptor (TNFRI). 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. MABTNFR1-B1 specifically binds to natural and recombinant truncated forms of TNFRI.

      552536 Rev. 2
      Format Details
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      Biotin
      Biotin is a ubiquitous co-factor (also known as Vitamin B7) that has many properties that make it extremely useful for molecular biology. Biotin has an extremely high affinity for the Avidin family of proteins (Kd = 10-15 M), making it the perfect tool to link two molecules. Biotin labeled antibodies can be combined with any number of Avidin-conjugated probes in order to customize an assay to a particular need. This is especially useful in the case of magnetic cell separation using streptavidin/magnetic bead conjugates, or in the case of flow cytometry using streptavidin/fluorophore conjugates.
      Biotin
      552536 Rev.2
      Citations & References
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      Development References (10)

      1. 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
      2. 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
      3. 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
      4. 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
      5. 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
      6. 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
      7. 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
      8. Munker R, DiPersio J, Koeffler HP. Tumor necrosis factor: receptors on hematopoietic cells. Blood. 1987; 70(6):1730-1734. (Biology). View Reference
      9. 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
      10. 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.
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      552536 Rev. 2

      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.

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