The production of IL-6 by 3T3 MEFs WT cultured with increasing concentrations of hIL-17A/F was assessed. Media from cells incubated with hIL-17A/F for 24 hours was collected and assayed for IL-6 by ELISA and the OD450 was determined.
The purity of recombinant hIL-17A/F was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hIL-17A/F and staining overnight with Coomassie Blue.
With carrier: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2 containing 20 μg BSA per 1 μg hIL-17A/F. Carrier free: Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.2.
Stable in lyophilized state at -20°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-17A/F. All lots are greater than 98% pure.
Less than 0.01 ng endotoxin/1 μg hIL-17A/F.
The bioactivity of recombinant hIL-17 A/F was determined by its ability to induce mouse IL-6 production by 3T3 MEFs WT. The ED50 of each lot is between 50-200 ng/ml.
Based on amino acid sequencing, greater than 80% of recombinant hIL-17A/F starts at Gly24 (GITIP) and the remainder starts at Ile20 (IVKAG). The non-glycosylated heterodimer has a calculated MW of 31,396. The DTT-reduced and non-reduced protein migrate as 38 kDa polypeptides. Lower mobility in SDS-PAGE is due to glycosylation.
Recombinant human IL-17A/F (hIL-17A/F) is a heterodimer that is composed of hIL-17A Ile20-Ala155 (Accession# NP_002181) linked to hIL-17F Arg31-Glu163 (Accession# NP_443104) via the linker GGGSGGGSGGGSGGGS. hIL-17A/F was expressed in human 293 cells at Cell Signaling Technology.
IL-17A/F is a cystine-linked heterodimer of IL-17A and IL-17F (1,2). IL-17A/F is produced by Th17 cells, and γδ T cells (1,2). IL-17A/F stimulates the production of pro-inflammatory cytokines and neutrophil chemoattractants, thereby functioning as a bridge between adaptive and innate immunity (1-3). Some research studies suggest that IL-17A/F may be involved in mucosal immunity against some bacterial infections and has a putative role in some autoimmune disorders (3,4). The receptor for IL-17A/F consists of a heterodimer of IL-17RA and IL-17RC and signaling through this receptor leads to the activation of the Erk1/2 and NF-κB pathways (1).
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