The proliferation of CTLL-2 cells treated with increasing concentrations of hIL-2 was assessed. After 48 hours treatment with hIL-2, cells were incubated with a tetrazolium salt and the OD450 - OD650 was determined.
The purity of recombinant hIL-2 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hIL-2 and staining overnight with Coomassie Blue.
Western blot analysis of extracts from CTLL-2 cells, untreated or treated with hIL-2 for 20 minutes, using Phospho-S6 Ribosomal Protein (Thr235/236)(2F9) Rabbit mAb #4856 (upper) or S6 Ribosomal Protein (5G10) Rabbit mAb #2217 (lower).
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hIL-2. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.
Stable in lyophilized state at 4°C for 1 year after receipt. Sterile stock solutions reconstituted with carrier protein are stable at 4°C for 2 months and at -20°C for 6 months. Avoid repeated freeze-thaw cycles.Maintain sterility. Storage at -20°C should be in a manual defrost freezer.
>98% as determined by SDS-PAGE of 6 μg reduced (+) and non-reduced (-) recombinant hIL-2. All lots are greater than 98% pure.
Recombinant hIL-2 does not have a Met on the amino terminus and has a calculated MW of 15,418. DTT-reduced and non-reduced protein migrate as 14 kDa polypeptides with non-reduced protein having slightly greater mobility due to an intramolecular cystine. The expected amino-terminal APTSS of recombinant hIL-2 was verified by amino acid sequencing.
The bioactivity of recombinant hIL-2 was determined in a CTLL-2 cell proliferation assay. The ED50 of each lot is between 0.1 - 1.0 ng/ml.
Less than 0.01 ng endotoxin/1 μg hIL-2.
Recombinant human IL-2 (hIL-2) Ala21-Thr153 (Accession #NM_000586) was produced in E. coli at Cell Signaling Technology.
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 (4, 5). 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 (3). 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-2 Rα 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).
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