Each lot of this antibody is quality control tested by immunofluorescent staining with flow cytometric analysis. The purified MP4-25D2 has been tested by blocking fluorochrome conjugated MP4-25D2 for intracellular cytokine staining. In order to obtain complete blocking results, a saturated amount of purified antibody (≤ 5.0 ug/million cells) should be used for incubation with target cells, prior to staining with fluorochrome conjugated antibody. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
ELISA1,3 or ELISPOT4,5 Detection: The biotinylated MP4-25D2 antibody is useful as a detection antibody for a sandwich ELISA or ELISPOT assay, when used in conjunction with purified 8D4-8 antibody (Cat. No. 500702/500707) as the capture antibody. Biotin-F(ab') (Cat. No. 500816) is recommended for ELISA analysis of human serum and plasma samples, to prevent non-specific binding of interfering substances such as rheumatoid factor and heterophilic antibodies. Each lot of this antibody is quality control tested by ELISA assay. Flow Cytometry6,9: The fluorochrome-labeled MP4-25D2 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify IL-4 -producing cells within mixed cell populations. View intracellular cytokine staining protocol Neutralization1-3: The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for neutralization of human IL-4 bioactivity (Cat. No. 500815).
Application References:
1. Chretien, I., et al., 1989. J. Immunol. Meth. 117: 67. 2. Ramanathan, L., et al., 1993. Biochem. 32: 3549. 3. Abrams, J., et al., 1992. Immunol. Rev. 127: 5. 4. Mahanty, S., et al., 1992. J. Immunol. 148: 3567. 5. Klinman, D., et al., 1994. Curr. Prot. Immunol.. John Wiley and Sons, New York. Unit 6.19. 6. Prussin, C., et al., 1995. J. Immunol. Meth. 188: 117. 7. Raqib, R., et al., 1995. Infect. Immun. 63: 289. 8. Andersson, J., et al., 1994. Immunology 83: 16. 9. Iwamoto, S., et al., 2007. J. Immunol. 179:1449. (ICFC) PubMed 10. Kubota, M. et al. 1997. J. Immunol. 158:5321. 11. Dzhagalov, I.,et al.2007. J. Immunol178:2113.PubMed
PMA/Ionomycin-stimulated human PBMCs were stained with CD3 PE/Cy5 and MP4-25D2 PE
Description:
IL-4 is a pleiotropic cytokine that is produced by activated T cells mast cells and basophils. IL-4 elicits many different biological responses, but has two dominant functions. The first is regulating differentiation of naïve CD4+ T cell to the Th2 type. Th2 cells produce IL-4, IL-5, IL-10 and IL-13, which tend to favor a humoral immune response while suppressing a cell-mediated immune response controlled by Th1 cells. The second is regulating IgE and IgG1 production by B cells. The MP4-25D2 antibody can neutralize the bioactivity of natural or recombinant IL-4.
Upregulated by IL-2, platelet activating factor; downregulated by TGF-β
Cellular Sources:
Mast cells, T cells, bone marrow stromal cells
Cellular Targets:
B cells, T cells, monocytes, endothelial cells, fibroblasts
Receptors:
Heterodimer IL-4Rα (CD124); γ-subunit (CD132) in common with IL-2R, IL-7R, IL-13R, IL-15R
Bioactivity/Activities:
Differentiation of naïve CD4+ T cells to the TH2 type, proliferation/differentiation of activated B cells, expression of class II MHC antigens, and of low affinity IgE receptors in resting B cells
Antigen References:
1. Fitzgerald, K., et al., Eds. 2001. The Cytokine FactsBook. Academic Press, San Diego. 2. Boulay, J., et al., 1992. Curr. Opin. Immunol. 4:294. 3. Dullens, H., et al., 1991. In vivo 5:567. 4. Paul, W. 1991. Blood. 77:1859.
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