Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Human Interleukin-5 (hIL-5) #5924

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Source

Recombinant human IL-5 (hIL-5) Ile20-Ser134 (Accession #NP_000870) was expressed in human 293 cells at Cell Signaling Technology.

Molecular Characterization

Recombinant hIL-5 contains no "tags" and the nonglycosylated protein has a calculated MW of 13,149. DTT-reduced protein migrates as a 14-21 kDa polypeptide. Heterogeneity and lower mobility in SDS-PAGE is due to glycosylation. The non-reduced cystine-linked homodimer migrates as a 32-38 kDa protein. The expected amino-terminal IPTEI of recombinant hIL-5 was verified by amino acid sequencing.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hIL-5. All lots are greater than 98% pure

Bioactivity

The activity of hIL-5 was assessed by quantification of phospho-p44/42 MAPK (Erk1/2) in butyric acid differentiated HL-60 Clone 15 cells. The observed ED50 values are between 0.50-1 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant hIL-5 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hIL-5 and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

The induction of phospho-p44/42 MAPK (Erk1/2) in butyric acid differentiated HL-60 Clone 15 cells in response to hIL-5. HL-60 Clone 15 cells were differentiated into eosinophil-like cells with butyric acid for 5 d. Cells were then treated with hIL-5 for 15 min, lysed, and p44/42 MAPK (Erk1/2) phosphorylation was quantified using the PathScan® Phospho-p44/42 MAPK (Thr202/Tyr204) Sandwich ELISA Kit #7177.

Western Blotting

Western Blotting

The induction of phospho-p44/42 MAPK (Erk1/2) in butyric acid differentiated HL-60 Clone 15 cells in response to hIL-5. HL-60 Clone 15 cells were differentiated into eosinophil-like cells with butyric acid for 5 d. Cells were then treated with hIL-5 for 15 min, lysed, and p44/42 MAPK (Erk1/2) phosphorylation was assessd by western blot using Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 and p44/42 MAPK (Erk1/2) (137F5) Rabbit mAb #4695.


Endotoxin

Less than 0.01 ng endotoxin/1 μg hIL-5.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hIL-5. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.

Background

IL-5 is a pleiotrophic cytokine that is predominantly produced by TH2 T cells but can also be expressed by activated eosinophils, mast cells, NK cells, and iNKT cells (1-4). Both human and mouse IL-5 are glycosylated disulfide-linked homodimers (1). The IL-5 receptor is a heterodimer that consists of a high affinity IL-5 binding α chain and the common β chain, which is shared by GM-CSF and IL-3 receptors, for signal transduction (1,2). Soluble IL-5Rα binds with high affinity to IL-5, thereby inhibiting IL-5 activity (1). IL-5-mediated signaling can activate the Erk1/2, Jak2, and Stat5 signaling pathways (1,2). In mice, IL-5 is important for the differentiation of antibody secreting cells from activated B cells (2). IL-5 induces eosinophil activation, proliferation, and differentiation in both mice and humans (2,3).

  1. Takatsu, K. (2011) Proc Jpn Acad Ser B Phys Biol Sci 87, 463-85.
  2. Wang, W. et al. (2011) PLoS One 6, e17766.
  3. Kouro, T. and Takatsu, K. (2009) Int Immunol 21, 1303-9.
  4. Chuang, Y.H. et al. (2011) J Immunol 186, 4687-92.

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For Research Use Only. Not For Use In Diagnostic Procedures.

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