Search functionality is temporarily unavailable due to a system issue. 5:30 p.m - 10:30 p.m CEST on Sunday, January 18, 2026.
Read More
If you encounter problems while ordering, please contact us at BDBCustomerService@bd.com
Expression of IL-4 by stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC were stimulated with soluble anti-human CD3 antibody (1 µg/ml final concentration; Cat. No. 555329), recombinant human IL-2 (10 ng/ml final concentration; Cat. No. 554603) and recombinant human IL-4 (10 ng/ml final concentration; Cat. No. 554605) for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days.
Finally, the cells were harvested and stimulated for 6 hours with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The cells were harvested, stained with PE-Cy5™ (formerly known as Cy-Chrome™) -anti CD4 (Cat. No. 555348), fixed, permeabilized, and subsequently
stained with 0.05 µg of PE-rat anti-human IL-4 antibody (PE-MP4-25D2, Cat. No. 554485) by using PharMingen's staining
protocol (left panel). To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabelled MP4-25D2 antibody (2.5 µg, Cat. No. 554482; right panel) prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabelled antibody blocking controls.
Expression of IL-4 by stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC were stimulated with soluble anti-human CD3 antibody (1 µg/ml final concentration; Cat. No. 555329), recombinant human IL-2 (10 ng/ml final concentration; Cat. No. 554603) and recombinant human IL-4 (10 ng/ml final concentration; Cat. No. 554605) for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days.
Finally, the cells were harvested and stimulated for 6 hours with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The cells were harvested, stained with PE-Cy5™ (formerly known as Cy-Chrome™) -anti CD4 (Cat. No. 555348), fixed, permeabilized, and subsequently
stained with 0.05 µg of PE-rat anti-human IL-4 antibody (PE-MP4-25D2, Cat. No. 554485) by using PharMingen's staining
protocol (left panel). To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabelled MP4-25D2 antibody (2.5 µg, Cat. No. 554482; right panel) prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabelled antibody blocking controls.
Expression of IL-4 by stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC were stimulated with soluble anti-human CD3 antibody (1 µg/ml final concentration; Cat. No. 555329), recombinant human IL-2 (10 ng/ml final concentration; Cat. No. 554603) and recombinant human IL-4 (10 ng/ml final concentration; Cat. No. 554605) for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days.
Finally, the cells were harvested and stimulated for 6 hours with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The cells were harvested, stained with PE-Cy5™ (formerly known as Cy-Chrome™) -anti CD4 (Cat. No. 555348), fixed, permeabilized, and subsequently
stained with 0.05 µg of PE-rat anti-human IL-4 antibody (PE-MP4-25D2, Cat. No. 554485) by using PharMingen's staining
protocol (left panel). To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabelled MP4-25D2 antibody (2.5 µg, Cat. No. 554482; right panel) prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabelled antibody blocking controls.
Expression of IL-4 by stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC were stimulated with soluble anti-human CD3 antibody (1 µg/ml final concentration; Cat. No. 555329), recombinant human IL-2 (10 ng/ml final concentration; Cat. No. 554603) and recombinant human IL-4 (10 ng/ml final concentration; Cat. No. 554605) for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days.
Finally, the cells were harvested and stimulated for 6 hours with PMA (Sigma, Cat. #P-8139) and calcium ionophore A23187 (Sigma) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The cells were harvested, stained with PE-Cy5™ (formerly known as Cy-Chrome™) -anti CD4 (Cat. No. 555348), fixed, permeabilized, and subsequently
stained with 0.05 µg of PE-rat anti-human IL-4 antibody (PE-MP4-25D2, Cat. No. 554485) by using PharMingen's staining
protocol (left panel). To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabelled MP4-25D2 antibody (2.5 µg, Cat. No. 554482; right panel) prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabelled antibody blocking controls.
The PE-MP4-25D2 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-4 producing cells within mixed cell populations (see image, left panel). For optimal immunofluorescent staining for flow cytometric analysis, this antibody should be titrated (≤ 0.20 µg mAb/million cells). For specific methodology, please visit the protocols section or chapter on Intracellular Staining in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. A useful control for demonstrating specificity of staining is the following: pre-block the fixed/permeabilized cells with unlabelled MP4-25D2 antibody (Cat. No. 554482) prior to staining. The intracellular cytokine staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized mouse and human cells is PE-R3-34 immunoglobulin (Cat. No. 554685); use at comparable concentrations to antibody of interest.
OTHER APPLICATIONS
ELISA Detection: The biotinylated MP4-25D2 antibody (Cat. No. 554483) is useful as a detection antibody for a sandwich ELISA for measuring human IL-4 protein levels. For testing IL-4 in serum or plasma, the BD OptEIA™ Human IL-4 ELISA set (Cat. No. 555194) or BD OptEIA Human IL-4 ELISA Kit (Cat. No. 550614) is recommended.
Neutralization: The NA/LE™ formatted MP4-25D2 antibody (Cat. No. 554481) has been reported to be useful for neutralization of human IL-4 bioactivity.
The MP4-25D2 monoclonal antibody specifically binds to Interleukin-4 (IL-4). IL-4 is also known as Lymphocyte stimulatory factor 1, B cell stimulatory factor 1 (BSF-1), or B cell growth factor 1 (BCGF-1). IL-4 is produced by activated T cells, basophils, and mast cells. IL-4 is a multifunctional cytokine and growth factor that affects a variety of different target cell types. IL-4 can costimulate T cell proliferation and survival, as well as help drive Th2-like cell differentiation and effector functions. It costimulates B cell proliferation and survival, and can help B cells differentiate into IgG4- or IgE-producing cells. IL-4 can also diminish the proinflammatory functions of monocytes and macrophages. IL-4 exerts its biological effects by binding with high affinity to cell surface CD124 (IL-4Rα chain). CD124 forms signaling IL-4 receptor complexes with either CD132/common γ chain or CD213a1/IL-13Rα1 to form either Type I or Type II IL-4 Receptor complexes, respectively. The immunogen used to generate the MP4-25D2 hybridoma was purified recombinant human IL-4. This is a neutralizing antibody. The MP4-25D2 antibody has been reported to crossreact with IL-4 from rhesus monkeys. The use of the MP4-25D2 antibody for epitope mapping of human IL-4 has been described.
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.
