The ability of Mouse IL-3 Neutralizing (D6C1) Rabbit mAb to inhibit mIL-3-induced BaF3 cell proliferation was assessed. Cells were incubated with increasing concentrations of antibody in the presence of mIL-3 #8923 (250 pg/ml). After 72 hr, viable cells were detected by incubation with a tetrazolium salt and the OD450 was determined.
The proliferation of BaF3 cells treated with increasing amounts of mIL-3 #8923 was determined. After a 72 hr treatment, cells were incubated with a tetrazolium salt and the OD450 was determined.
Neutralizing antibodies can be used to inhibit normal biological function through their binding to biological molecules. These reagents can be used to determine the effects that a particular molecule has in biological systems. Mouse IL-3 Neutralizing (D6C1) Rabbit mAb has been shown to neutralize the proliferation of IL-3-dependent BaF3 cells in vitro with an ND50 in the range of 0.5-3.5 µg/ml.
CST recommends incubation of the neutralizing antibody with the intended target for 1 hr at 37ºC before addition to the experiment at an optimal concentration determined by the user.
Add sterile 10 mM HEPES pH 7.0 to a final concentration of greater than 50 μg/ml. Solubilize for 20 min at room temperature with occasional gentle vortexing.
Lyophilized from a 0.2 µm filtered solution in 10 mM HEPES with trehalose.
Store lyophilized material at -20ºC. After reconstitution, recommended storage at 4ºC for 1 month or -20ºC for 6 months. Avoid repeated freeze/thawing.
Mouse IL-3 Neutralizing (D6C1) Rabbit mAb binds to mouse IL-3 and neutralizes its effects in a BaF3 cell proliferation assay. This antibody does not cross-react with human IL-3, human IL-5, human GM-CSF, or mouse GM-CSF.
Monoclonal antibody is produced by immunizing animals with a recombinant mouse IL-3 protein.
IL-3 is produced by T cells, mast cells, and eosinophils (1). Target cells include hematopoietic progenitors, neutrophils, macrophages, mast cells, eosinophils, lymphoid, and erythroid cells (1). IL-3 supports growth and differentiation and is used as a media additive to support the culture of many cell types (1). The IL-3 receptor is a heterodimer of the IL-3-specific α-chain and the common β-chain, βc, which is also used by GM-CSF and IL-5. (1). Binding of IL-3 can also involve substitution of βc by a βIL-3-chain that appears to be specific for IL-3 (1,2). Binding of IL-3 to its cognate receptor(s) induces activation of Jak2 and the PI3K/Akt pathway, and phosphorylation of Stat1/3/5/6 (1). IL-3 may play an important role in the development of airway inflammation associated with asthma (3-5).
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