Cell Signaling Technology

Product Pathways - Growth Factors/Cytokines

Mouse Interleukin-2 (mIL-2) #5201

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Source

Recombinant mouse IL-2 (mIL-2) Ala21-Gln169 (Accession #NP_032392) was produced in E. coli at Cell Signaling Technology.

Molecular Characterization

Based on amino acid sequencing, greater than 50% of recombinant mIL-2 has a Met on the amino-terminal Ala21 (MAPTS) and has a calculated MW of 17,231. The remainder starts at Ala21 (APTSS) or Pro22 (PTSSP). DTT-reduced and non-reduced protein migrate as 19 kDa polypeptides.

Purity

>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant mIL-2. All lots are greater than 98% pure.

Bioactivity

The bioactivity of recombinant mIL-2 was determined in a CTLL-2 cell proliferation assay. The ED50 of each lot is between 0.5-2 ng/ml.

Coomassie Gel

Coomassie Gel

The purity of recombinant mIL-2 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant mIL-2 and staining overnight with Coomassie Blue.

Bioactivity

Bioactivity

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

Western Blotting

Western Blotting

Western blot analysis of extracts from CTLL-2 cells untreated or treated with mIL-2 for 10 min, using Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb #9359 (upper) and Stat5 (3H7) Rabbit mAb #9358 (lower).


Endotoxin

Less than 0.01 ng endotoxin/1 μg mIL-2.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 1 mM DTT and 20 μg BSA per 1 μg mIL-2. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 1 mM DTT.

Background

IL-2 is a T cell stimulatory cytokine best known for inducing T cell proliferation and NK cell proliferation and activation (1,2). IL-2 also promotes peripheral development of Treg cells (3,4). Conversely, IL-2 is involved in activation induced cell death (AICD) that is observed post T cell expansion by increasing levels of Fas on CD4+ T cells (5). The effects of IL-2 are mediated through a trimeric receptor complex consisting of IL-2Rα, IL-2Rβ, and the common gamma chain, γc (1,2). IL-2Rα binds exclusively to IL-2 with low affinity and increases binding affinity of the whole receptor complex including IL-2Rβ and γc subunits. IL-2Rβ is also used by IL-15 (1,2). The common γc is used by other cytokines including IL-4, IL-7, IL-9, IL-15, and IL-21 (1,2). Binding of IL-2 initiates signaling cascades involving Jak1, Jak3, Stat5, and the PI3K/Akt pathways (1,2).

  1. Ma, A. et al. (2006) Annu Rev Immunol 24, 657-79.
  2. Gaffen, S.L. and Liu, K.D. (2004) Cytokine 28, 109-23.
  3. Fehérvari, Z. et al. (2006) Trends Immunol 27, 109-11.
  4. Antony, P.A. et al. (2006) J Immunol 176, 5255-66.
  5. Jaleco, S. et al. (2003) J Immunol 171, 61-8.

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