Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Mouse Interleukin-4 (mIL-4) #5208

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Source

Recombinant mouse IL-4 (mIL-4) His21-Ser140 (Accession #NP_067258) was expressed in human 293 cells at Cell Signaling Technology.

Molecular Characterization

Recombinant mIL-4 contains no "tags" and the nonglycosylated protein has a calculated MW of 13,557. DTT-reduced and non-reduced protein migrate as 16 kDa polypeptide due to glycosylation. The expected amino-terminal HIHGC of recombinant mIL-4 was verified by amino acid sequencing.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mIL-4. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant mIL-4 was determined in an HT-2 cell proliferation assay. The ED50 of each lot is between 100-400 pg/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant mIL-4 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mIL-4 and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The proliferation of HT-2 cells treated with increasing concentrations of mIL-4 was assessed. After 48 hour treatment with mIL-4, cells were incubated with a tetrazolium salt and the OD450 - OD650 was determined.

Western Blotting

Western Blotting

Western blot analysis of extracts from HT-2 cells untreated or treated with mIL-4 for 10 minutes, using Phospho-Jak1 (Tyr1022/1023) Antibody #3331 (upper) or Jak1(6G4) Rabbit mAb #3344 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1 μg mIL-4.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg mIL-4. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.

Background

IL-4 is produced by T cells, NK T cells, γδ cells, and mast cells (1). Target cells include B cells, T cells, and macrophages (1). IL-4 induces the polarization of naïve helper T cells into the TH2 phenotype (1,2). IL-4 also promotes B cell proliferation, antibody class switching and the production other TH2 cytokines including IL-5 and IL-9. IL-4 induced TH2 polarization is important in developing humoral immunity against extracellular pathogens (1) and is involved in the development of allergy and asthma (3). IL-4 binds to two distinct receptors, the Type I receptor and Type II receptor. Type I receptor is a heterodimer consisting of IL-4Rα chain and the common gamma chain, γc (4,5). Type II receptor, which is shared with IL-13, is a heterodimer of IL-4Rα and IL-13Rα1. Signaling initiated via Type I receptor results in the activation of Jak1/Stat6, Jak3 and the PI3K/Akt pathways (4). The Type II receptor activates the Jak1/Stat6 and the Tyk2/Stat3 pathways (4).

  1. Corthay, A. (2006) Scand J Immunol 64, 93-6.
  2. Wynn, T.A. (2003) Annu Rev Immunol 21, 425-56.
  3. Nakajima, H. and Takatsu, K. (2007) Int Arch Allergy Immunol 142, 265-73.
  4. Wills-Karp, M. and Finkelman, F.D. (2008) Sci Signal 1, pe55.
  5. Mueller, T.D. et al. (2002) Biochim Biophys Acta 1592, 237-50.

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