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准备和存储
推荐的实验流程
ELISA Capture: The purified 55-3 antibody (Cat. No. 551403) is useful as a capture antibody for a sandwich ELISA for measuring human soluble CD14 protein levels. Purified 55-3 antibody can be paired with the biotinylated 3-C39 anti-human sCD14 (Cat. No. 551405) as the detecting antibody, with recombinant soluble CD14 as the standard. Purified 55-3 antibody should be titrated between 1-4 µg/ml to determine its optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of recombinant human soluble CD14, ranging from 2000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on ELISA in the Immune Function Handbook.
商品通知
- 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 55-3 monoclonal antibody reacts with human CD14. CD14 is a 53 - 55 kDa cell surface glycoprotein that is expressed by monocytes, macrophages, and activated granulocytes as a glycophosphatidylinositol-linked protein. CD14 functions as a receptor for lipopolysaccharide (LPS, endotoxin) when LPS is bound to LBP (LPS binding protein). Soluble forms of CD14 (sCD14) can be detected either in serum or in supernatants from cell cultures. Soluble CD14 is produced either by shedding of membrane CD14 or by release from cells before addition of the GPI anchor. Both membrane and sCD14 function to enhance cell activation by LPS. Binding of LPS to CD14 activates monocytes or macrophages to produce cytokines and express adhesive proteins and low molecular weight proinflammatory mediators. Non-CD14 expressing cells, such as endothelial cells, can be activated by a complex of LPS and sCD14. Serum levels of sCD14 increase in association with sepsis, infectious disease, autoimmunity, and allergic disorders. The gene for human CD14 has been cloned and sequenced and recombinant sCD14 has been shown to be protective against LPS-induced lethality. The immunogen used to generate the 55-3 hybridoma was purified, soluble recombinant human CD14 protein.
研发参考 (10)
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Arditi M, Zhou J, Dorio R, Rong GW, Goyert SM, Kim KS. Endotoxin-mediated endothelial cell injury and activation: role of soluble CD14. Infect Immun. 1993; 61(8):3149-3156. (Biology). 查看参考
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Egerer K, Feist E, Rohr U, Pruss A, Burmester GR, Dorner T. Increased serum soluble CD14, ICAM-1 and E-selectin correlate with disease activity and prognosis in systemic lupus erythematosus. Lupus. 2000; 9(8):614-621. (Biology). 查看参考
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Ferrero E, Goyert SM. Nucleotide sequence of the gene encoding the monocyte differentiation antigen, CD14. Nucleic Acids Res. 1988; 16(9):4173. (Clone-specific). 查看参考
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Goyert SM, Ferrero E, Rettig WJ, Yenamandra AK, Obata F, Le Beau MM. The CD14 monocyte differentiation antigen maps to a region encoding growth factors and receptors. Science. 1988; 239(4839):497-500. (Biology). 查看参考
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Goyert SM, Lohen L, Gangloff SC, Ashmun R, Haeffner Cavaillon N. CD14 Workshop Panel report. In: Kishimoto T. Tadamitsu Kishimoto .. et al., ed. Leucocyte typing VI : white cell differentiation antigens : proceedings of the sixth international workshop and conference held in Kobe, Japan, 10-14 November 1996. New York: Garland Pub.; 1997:36.
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Haziot A, Chen S, Ferrero E, Low MG, Silber R, Goyert SM. The monocyte differentiation antigen, CD14, is anchored to the cell membrane by a phosphatidylinositol linkage. J Immunol. 1988; 141(2):547-552. (Clone-specific). 查看参考
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Haziot A, Rong GW, Lin XY, Silver J, Goyert SM. Recombinant soluble CD14 prevents mortality in mice treated with endotoxin (lipopolysaccharide). J Immunol. 1995; 154(12):6529-6532. (Clone-specific). 查看参考
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Haziot A, Rong GW, Silver J, Goyert SM. Recombinant soluble CD14 mediates the activation of endothelial cells by lipopolysaccharide. J Immunol. 1993; 151(3):1500-1507. (Biology). 查看参考
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Nockher WA, Wick M, Pfister HW. Cerebrospinal fluid levels of soluble CD14 in inflammatory and non-inflammatory diseases of the CNS: upregulation during bacterial infections and viral meningitis. J Neuroimmunol. 1999; 101(2):161-169. (Biology). 查看参考
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Ulevitch RJ, Tobias PS. Receptor-dependent mechanisms of cell stimulation by bacterial endotoxin. Annu Rev Immunol. 1995; 13:437-457. (Biology). 查看参考
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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