TARC production from human PBMC treated with increasing concentrations of hHis6TSLP was assessed. After 24 hours, cell supernatants were harvested and assayed for TARC by ELISA and the OD450-OD650 was determined.
The purity of recombinant hHis6TSLP was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hHisTSLP 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 hHis6TSLP. 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 hHis6TSLP. All lots are greater than 98% pure.
Recombinant human N-terminally His6-tagged TSLP has a calculated MW of 17,352. DTT-reduced and non-reduced protein migrate as 31 kDa polypeptides. Lower mobility in SDS-PAGE is due to glycosylation. The expected amino terminus of recombinant hHis6TSLP was verified by amino acid sequencing.
The bioactivity of recombinant hHis6TSLP was determined by its ability to induce TARC production by PBMC. The ED50 of each lot is between 50-200 pg/ml.
Less than 0.01 ng endotoxin/1 μg hHis6TSLP.
Recombinant human His6TSLP (hHis6TSLP) Tyr29-Gln159 (Accession #NP_149024) was expressed in human 293 cells at Cell Signaling Technology. Recombinant hHis6TSLP contains 2 point mutations to eliminate potential furin cleavage sites (R127A and R130S).
TSLP is an IL-7-like cytokine that plays a key role in promoting TH2–type immune responses (1). Epithelial cells in the skin, lung, and gut are the predominant producers of TSLP (1). A number of immune cell types are responsive to TSLP including dendritic cells, lymphocytes, mast cells, and basophils, among others (1). Consistent with its role as a promoter of TH2-type immune responses, TSLP is a crucial component of the anti-helminth immune response (1). Moreover, TSLP may promote tumor progression by inducing TH2-type inflammation (2-4). TSLP signals through IL-7Rα/TSLPR heterodimers, activating Jak1/2 and Stat5 pathways (5,6).
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