|45095||Human Interleukin-21 (hIL-21)||
The production of IFN-γ by NK-92 cells cultured with increasing concentrations of hIL-21 was assessed. Media from cells incubated with hIL-21 for 24 hours was collected and assayed for IFN-γ by ELISA and the OD450 was determined.
The purity of recombinant hIL-21 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hIL-21 and staining overnight with Coomassie Blue.
Western blot analysis of extracts from NK-92 cells, untreated or treated with hIL-21 for 15 minutes, using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145 (upper) or Stat3 Antibody #9132 (lower).
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hIL-21. 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-21. Less than 9% is nicked after Arg114 to generate 6 kDa and 10 kDa polypeptides observed in reduced (+) hIL-21.
Recombinant hIL-21 contains no "tags" and the nonglycosylated protein has a calculated MW of 15,463. DTT-reduced and non-reduced protein migrate as 16 kDa polypeptides, with non-reduced having slightly greater mobility due to intramolecular cystines. The expected amino-terminal QGQDR of recombinant hIL-21 was verified by amino acid sequencing.
The bioactivity of recombinant hIL-21 was determined by its ability to induce IFN-γ production by NK-92 cells. The ED50 of each lot is between 1.0-6.0 ng/ml.
Less than 0.01 ng endotoxin/1 μg hIL-21.
Recombinant human IL-21 (hIL-21) Gln30-Ser155 (Accession # NM_021803) was expressed in human 293 cells at Cell Signaling Technology.
IL-21 is produced by Th17, T follicular helper (Tfh) and NKT cells (1-3). Targets include T cells, B cells, NK cells and dendritic cells (1-3,5). IL-21 induces proliferation and activation of NK cells, thereby up-regulating IFN-γ production and cytotoxic activity (1,3,4). IL-21 increases proliferation and survival of CD40 primed B cells (1,2) and appears to have a significant role in plasma cell differentiation (2). IL-21 binds to a complex consisting of IL-21Rα and the common γ chain, γc. IL-21 binding activates the Jak1/Jak3, Stat1, Stat3 and Stat5 pathway. IL-21 binding can also activate the MAP kinase and PI3K/Akt pathways.
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