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Expression of cell surface TNFRII by whole-lysed human blood. Whole human blood was lysed with PharmLyse™ (Cat. No. 555899) prior to staining with hTNFR-M1. Whole lysed human blood was subsequently blocked with normal polyclonal human IgG and stained with purified hTNFR-M1 (0.06 µg/10e6 cells, Cat No. 551311) followed by biotinylated goat anti-rat F(ab')2 IgG and streptavidin phycoerythrin (0.015 µg, Cat. No. 554061). Staining with the hTNFR-M1 antibody (filled histograms) is compared to staining obtained using the isotype control antibody (open histograms). The histograms in the figure were derived from gated events with the light scattering characteristics of viable lymphocytes (left panel), monocytes (center panel) and granulocytes (right panel). 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.
BD Pharmingen™ Purified Rat Anti-Human CD120b
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준비 및 보관
권장 분석 절차
Immunofluorescent Staining and Flow Cytometric Analysis: The purified hTNFR-M1 (Cat. No. 551311) antibody can be used for the immunofluorescent staining (≤ 1 µg antibody/10e6 cells) and flow cytometric analysis of human nucleated cells to measure their expressed levels of surface TNFRII. An appropriate immunoglobulin isotype control is clone R35-38 (Cat. No. 555846). A three-layer staining protocol is recommended for maximizing the detection of TNFRII expressed by cells as detailed in the figure legend.Please note also that as a consequence of in vivo or in vitro activation, cell surface TNFRII 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 TNFRII.
제품 고시
- 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.
관련 제품
The hTNFR-M1 antibody specifically recognizes the extracellular domain of the 75 kDa transmembrane receptor for the human cytokines, tumor necrosis factor (TNF or TNF-α) and lymphotoxin-alpha (LT-α3, aka, lymphotoxin or TNF-β). This receptor is referred to as the p75 or Type II Tumor Necrosis Factor Receptor (TNFRII) [aka, CD120b]. Human TNFRII proteins are expressed by hematopoietic cells including macrophages, neutrophils, lymphocytes, thymocytes and mast cells. TNFRII is expressed by a variety of other cell types including endothelial cells, cardiac myocytes and prostate cells. Naive B cells express very low or undetectable levels of TNFRII whereas mature erythrocytes and platelets are uniformly negative for TNFRII expression. The immunogen used to generate the hTNFR-M1 hybridoma was COS- expressed recombinant human TNFRII.
개발 참고 자료 (11)
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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). 참조 보기
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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). 참조 보기
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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). 참조 보기
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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). 참조 보기
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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). 참조 보기
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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. (Clone-specific: Flow cytometry). 참조 보기
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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). 참조 보기
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Munker R, DiPersio J, Koeffler HP. Tumor necrosis factor: receptors on hematopoietic cells. Blood. 1987; 70(6):1730-1734. (Clone-specific: Flow cytometry). 참조 보기
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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). 참조 보기
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Ware CF, Crowe PD, Vanarsdale TL, et al. Tumor necrosis factor (TNF) receptor expression in T lymphocytes. Differential regulation of the type I TNF receptor during activation of resting and effector T cells.. J Immunol. 1991; 147(12):4229-38. (Clone-specific). 참조 보기
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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.
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