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The antibody was purified by affinity chromatography, and conjugated with PE/Cy7 under optimal conditions. The solution is free of unconjugated PE/Cy7 and unconjugated antibody.
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. Test size products are transitioning from 20 µl to 5 µl per test. Please check your vial or your CoA to find the suggested use of this reagent per million cells in 100 µl staining volume or per 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application. Read more at www.biolegend.com/testsize regarding the test size change.
COA:
Application Notes:
ELISA Detection1,3 or ELISPOT Detection4,5: 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. 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). The MP4-25D2 antibody can neutralize the bioactivity of natural or recombinant IL-4.
Cy3, Cy5, Cy5.5 and Cy7 are subject to proprietary rights of GE Healthcare Bio-Sciences Corp. and Carnegie Mellon University and made and sold under license from GE Healthcare Bio-Sciences Corp. Sale of this product is licensed for research use only.
Application References:
1. Chretien I, et al. 1989. J. Immunol. Methods 117:67. (ELISA Dectection, Neut) 2. Ramanathan L, et al. 1993. Biochem. 32:3549. (Neut) 3. Abrams J, et al. 1992. Immunol. Rev. 127:5. (ELISA Dectection, Neut) 4. Mahanty S, et al. 1992. J. Immunol. 148:3567. (ELISPOT Dectection) 5. Klinman D, et al. 1994. Curr. Prot. Immunol. John Wiley and Sons New York. Unit 6.19. (ELISPOT Dectection) 6. Prussin C, et al. 1995. J. Immunol. Methods 188:117. (ICFC) 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. Immunol. 178:2113. PubMed
PMA+ionomycin-stimulated (6 hours) human peripheral blood lymphocytes surface stained with CD3 FITC, then intracellularly stained with MP4-25D2 PE/Cy7 (top) or rat IgG1, k PE/Cy7 isotype control (bottom)
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.
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.
*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/terms). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
APC anti-human IL-4
PMA + ionomycin-stimulated (6 hours) human peripheral blood lymphocytes intracellular stained with MP4-25D2 APC and CD3 (UCHT1) PE
Biotin anti-human IL-4
FITC anti-human IL-4
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with MP4-25D2 FITC and CD3 (UCHT1) APC
PE anti-human IL-4
PMA/Ionomycin-stimulated human PBMCs were stained with CD3 PE/Cy5 and MP4-25D2 PE
Alexa Fluor® 488 anti-human IL-4
PMA+ionomycin-stimulated (6 hours) human peripheral blood lymphocytes intracellular stained with MP4-25D2 Alexa Fluor®
488 and CD4 (RPA-T4) PE
Alexa Fluor® 647 anti-human IL-4
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with rat IgG1 Alexa Fluor® 647 isotype control and CD3 (UCHT1) PE
PMA/ionomycin-stimulated (6 hours) human peripheral blood lymphocytes stained with MP4-25D2 Alexa Fluor® 647 isotype control and CD3 (UCHT1) PE
Brilliant Violet 421™ anti-human IL-4
Human peripheral blood lymphocytes were stimulated with PMA + ionomycin for 6 hours (in the presence of monensin), surface stained with CD3 FITC, fixed, permeabilized and then stained with IL-4 (clone MP4-25D2) Brilliant Violet 421™ (top) or rat IgG1, κ Brilliant Violet 421™ isotype control (bottom).
PerCP/Cy5.5 anti-human IL-4
PMA/ionmycin-stimulated (6 hours) human peripheral blood lymphocytes intracellularly stained with CD3 FITC and MP4-25D2 PerCP/Cy5.5
PE/Cy7 anti-human IL-4
PMA+ionomycin-stimulated (6 hours) human peripheral blood lymphocytes surface stained with CD3 FITC, then intracellularly stained with MP4-25D2 PE/Cy7 (top) or rat IgG1, k PE/Cy7 isotype control (bottom)
