Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PS00-67] to IL-4 (Capture) (HA721230).
Target
Function
Cytokine secreted primarily by mast cells, T-cells, eosinophils, and basophils that plays a role in regulating antibody production, hematopoiesis and inflammation, and the development of effector T-cell responses. Induces the expression of class II MHC molecules on resting B-cells. Enhances both secretion and cell surface expression of IgE and IgG1. Regulates also the expression of the low affinity Fc receptor for IgE (CD23) on both lymphocytes and monocytes. Positively regulates IL31RA expression in macrophages. Stimulates autophagy in dendritic cells by interfering with mTORC1 signaling and through the induction of RUFY4. In addition, plays a critical role in higher functions of the normal brain, such as memory and learning. Upon binding to IL4, IL4R receptor dimerizes either with the common IL2R gamma chain/IL2RG to produce the type 1 signaling complex, located mainly on hematopoietic cells, or with the IL13RA1 to produce the type 2 complex, which is expressed also on nonhematopoietic cells. Engagement of both types of receptors initiates JAK3 and to a lower extend JAK1 phosphorylation leading to activation of the signal transducer and activator of transcription 6/STAT6.
Background References
1. Carr C., Aykent S., Kimack N.M., Levine A.D. Disulfide assignments in recombinant mouse and human interleukin 4. Biochemistry 30:1515-1523 (1991)
2. LaPorte S.L., Juo Z.S., Vaclavikova J., Colf L.A., Qi X., Heller N.M., Keegan A.D., Garcia K.C. Molecular and structural basis of cytokine receptor pleiotropy in the interleukin-4/13 system. Cell 132:259-272 (2008)
Sandwich ELISA analysis of IL-4 matched pair antibodies Elisa assay was performed by coating wells of a 96-well plate with 50 µl per well of capture antibody HA721230 [PS00-67] diluted in carbonate/bicarbonate buffer, at a concentration of 1 µg/mL overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 1%BSA/PBST blocking buffer, and incubated with serial diluted Recombinant IL-4 protein starting from 2000 pg/ml to 30.5 pg/ml for 1 hour at 37℃. The plate was washed and incubated with 50 µl per well of detect antibody HA721368 [PS00-66] (Biotin, 1:4000) for 1 hour at 37℃. Then the plate was washed and incubated with 50 µl per well of Streptavidin-HRP for 0.5 hour at 37℃. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.
Capture/detect antibodies, Recombinant IL-4 protein, Streptavidin-HRP are diluted in 1% BSA/PBST.
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