준비 및 보관
권장 분석 절차
ELISA Capture: For a mouse IL-5 ELISA, purified TRFK5 antibody can be paired with the biotinylated TRFK4 anti-mouse IL-5 antibody (Cat. No. 554397) as the detecting antibody, with recombinant mouse IL-5 (Cat. No. 554581) as the standard. For a human IL-5 ELISA, purified TRFK5 antibody can be paired with the biotinylated JES1-5A10 anti-human IL-5 antibody (Cat. No. 554491) as the detecting antibody, with recombinant human IL-5 (Cat. No. 554606) as the standard. The purified TRFK5 antibody should be titrated 1.0 - 4.0 µg/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of IL-5 protein ranging ~2000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit the protocols section on our website at http://www.bdbiosciences.com/support/resources/elisa_elispot/.
Note: For testing IL-5 in serum or plasma, the Mouse IL-5 OptEIA™ Set (Cat. No. 555236) and the Human IL-5 OptEIA™ Set (Cat. No. 555202) are recommended.
Immunofluorescent Staining and Flow Cytometric Analysis: The TRFK5 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate mouse IL-5 producing cells within mixed cell populations. PE- or APC-conjugated TRFK5 antibodies (Cat. No. 554395, Cat. No. 554396) are especially suitable for these experiments.
Neutralization: The BD Pharmingen™ NA/LE TRFK5 antibody (Cat. No. 554391) is useful for neutralization of mouse and human IL-5 bioactivity.
WB: The TRFK5 antibody has been found useful for Western blotting
제품 고시
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
관련 제품
The TRFK5 antibody reacts with mouse interleukin-5 (IL-5) and cross-reacts with human IL-5. The TRFK5 antibody has been reported to cross react with IL-5 from rhesus monkey. This is a neutralizing antibody.
개발 참고 자료 (9)
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Abrams J. Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies. Curr Protoc Immunol. 2001; 1:6.20-6.21. (Clone-specific: ELISA). 참조 보기
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Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Clone-specific: Neutralization). 참조 보기
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Assenmacher M, Schmitz J, Radbruch A. Flow cytometric determination of cytokines in activated murine T helper lymphocytes: expression of interleukin-10 in interferon-gamma and in interleukin-4-expressing cells. Eur J Immunol. 1994; 24(5):1097-1101. (Clone-specific: Flow cytometry). 참조 보기
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Mo XY, Sarawar SR, Doherty PC. Induction of cytokines in mice with parainfluenza pneumonia. J Virol. 1995; 69(2):1288-1291. (Clone-specific: ELISA). 참조 보기
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Sander B, Hoiden I, Andersson U, Moller E, Abrams JS. Similar frequencies and kinetics of cytokine producing cells in murine peripheral blood and spleen. Cytokine detection by immunoassay and intracellular immunostaining. J Immunol Methods. 1993; 166(2):201-214. (Clone-specific). 참조 보기
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Sarawar SR, Sangster M, Coffman RL, Doherty PC. Administration of anti-IFN-gamma antibody to beta 2-microglobulin-deficient mice delays influenza virus clearance but does not switch the response to a T helper cell 2 phenotype. J Immunol. 1994; 153(3):1246-1253. (Clone-specific: ELISA). 참조 보기
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Schumacher JH, O'Garra A, Shrader B, et al. The characterization of four monoclonal antibodies specific for mouse IL-5 and development of mouse and human IL-5 enzyme-linked immunosorbent. J Immunol. 1988; 141(5):1576-1581. (Clone-specific: Neutralization). 참조 보기
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Swain SL, Weinberg AD, English M, Huston G. IL-4 directs the development of Th2-like helper effectors. J Immunol. 1990; 145(11):3796-3806. (Clone-specific: Neutralization). 참조 보기
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Taguchi T, McGhee JR, Coffman RL, et al. Analysis of Th1 and Th2 cells in murine gut-associated tissues. Frequencies of CD4+ and CD8+ T cells that secrete IFN-gamma and IL-5. J Immunol. 1990; 145(1):68-77. (Clone-specific: ELISA). 참조 보기
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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.
