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Encompassing Amino Acids: 25-153(end)
Application: Useful for binding assays.
Assay Conditions: A white 96-well plate was precoated with IL-4Ra/CD124, Fc-Fusion (IgG1), Avi-Tag, Recombinant (#102467) (100 ng/well). Blocking Buffer 2 (#79728) and PBST were used for blocking and washing steps, respectively. For protein titration a serial dilution of IL-4, Avi-His-Tag, Biotin-Labeled Recombinant (#102539) at the highest concentration of 1 ng/µl (25 ng/reaction) was prepared in 1x PP-02 Assay Buffer. Diluted IL-4 was then added to the wells and incubated at Room Temperature (RT) for 1 hour. The plate was then washed again and incubated with Streptavidin-HRP conjugate diluted 1:1000 in Blocking Buffer 2 for 1 hour at RT. After the final wash, ELISA ECL substrate mix (#79670) was added to each well and chemiluminescence was read.
Background: Interleukin-4 is produced mainly by a subpopulation of activated T cells (Th2) that are the biologically most active helper cells for B cells and that also secrete IL-5 and IL-6. The biological activities of IL-4 are mediated by a specific receptor. The extracellular domain of the IL-4 receptor is related to the receptors for EPO (erythropoietin), IL-6, and the beta chain of the IL-2 receptor (CD122). There are two types of receptors for IL-4. Type 1 receptor is a heterodimer consisting of IL-4Ralpha (IL-4Rα) and IL-4Rgamma (IL-4Rγc), mostly present on hematopoietic cells. The type 2 receptor is a heterodimer consisting of IL-4Ralpha and IL-13Ralpha1 (Il-13Rα1). IL-4 enhances expression of MHC (major histocompatibility class) 2 antigens on B cells. It can promote their capacity to respond to other B cell stimuli and to present antigens for T cells. Pretreatment of macrophages with IL-4 prevents the production of IL-1, TNFα (tumor necrosis factor alpha) and prostaglandins in response to activation of the cells by bacterial endotoxins or IFN-γ (interferon gamma). IL-4 plays crucial roles in inflammation, fibrosis, allergic reactions, and cancer. The therapeutic exploitation of IL-4 signaling pathways can result in advances in the treatment of auto-immune disorders.
Description: Recombinant IL-4 (interleukin 4), transcript variant 1, encompassing amino acids 25-153(end). This construct contains a C-terminal Avi-Tag™ followed by a His-tag (6xHis). The recombinant protein was enzymatically biotinylated using the Avi-Tag™ and affinity purified.
Endotoxin Level: ≤11.5 EU/mg
Format: Aqueous buffer solution
Formulation: 8 mM phosphate, pH 7.4, 110 mM NaCl, 2.2 mM KCl, and 20% glycerol
Genbank: NM_000589.4
Host Cell Line: HEK293
Purity: ≥85%
Storage Stability: At least 6 months at –80°C.
Tags: Avi-His
Target: IL-4
Uniprot: P05112
Unit Definition: Protein
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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