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The antibody was purified by affinity chromatography, and conjugated with PE under optimal conditions. The solution is free of unconjugated PE and unconjugated antibody.
Concentration:
test sizes: lot-specific; µg size: 0.2 mg/ml
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:
ELISA3 or ELISPOT4 Capture: The purified 8D4 antibody is useful as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated MP4-25D2 antibody (Cat. No. 500804/500816) as the detecting antibody. The LEAF™ purified antibody is suggested for ELISPOT capture. Flow Cytometry2,5: The fluorochrome-labelled 8D4-8 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: 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. 500707). Additional reported applications (for the relevant formats) include: immunoprecipitation, immunohistochemical staining6 of paraformaldehyde-fixed, saponin-treated frozen tissue sections, and immunocytochemistry. Note: For testing human IL-4 in serum or plasma, BioLegend's ELISA Max™ Sets (Cat. No. 430301 to 430306) are specially developed and recommended.
Application References:
1. Bird, C., et al. 1991. Cytokine. 3:562. 2. Prussin, C., et al. 1995. J. Immunol. Meth. 188:117. 3. Hickie, I., et al. 2001. Twin Res. 4:94. 4. Heeger, P., et al. 1999. J. Immunol. 163:2267. 5. Jason, J., et al. 1999. Clin. Diagn. Lab Immunol. 6:73. 6. Andersson, U., et al. 1999. Detection and quantification of gene expression. New York:Springer-Verlag. 7. Gong, G., et al. 2008. Blood PubMed 8. Yoshino, N., et al. 2000. Exp. Anim. (Tokyo) 49:97. (FC)
Enriched human CD4+ T cells were stimulated with PMA+ionomycin, then surface stained with CD4 PE/Cy5 and intracellular stained with 8D4-8 PE
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 8D4-8 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.
*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.
PE anti-human IL-4
Enriched human CD4+ T cells were stimulated with PMA+ionomycin, then surface stained with CD4 PE/Cy5 and intracellular stained with 8D4-8 PE
LEAF™ Purified anti-human IL-4
Purified anti-human IL-4
APC anti-human IL-4
PMA+ionomycin-stimulated human peripheral blood mononuclear cells surface stained CD3 PE and intracellularly stained with 8D4-8 APC (top), bottom data demonstrates staining can be specifically blocked by pre-incubation of the cells with purified 8D4-8 antibody (gated on lymphocyte population).
Alexa Fluor® 488 anti-human IL-4
PMA+ionomycin-stimulated (6 hours) human peripheral blood lymphocytes surface stained with CD3 PE, then intracellular stained with 8D4-8 Alexa Fluor® 488 (top), bottom data shows the 8D4-8 Alexa Fluor® 488 staining can be specifically blocked by pre-incubation with purified 8D4-8 antibody.
Alexa Fluor® 647 anti-human IL-4
PMA+ionomycin-stimulated (6 hours) human peripheral blood lymphocytes surface stained with CD3 PE, then intracellular stained with 8D4-8 Alexa Fluor® 647 (top), bottom data shows the 8D4-8 Alexa Fluor® 647 staining can be specifically blocked by pre-incubation with purified 8D4-8 antibody
