The inhibition of IL-4 induced proliferation in HT-2 cells treated with increasing concentrations of hTGF-β2 was assessed. After 48 hour treatment with hTGF-β2, cells were incubated with a tetrazolium salt and the OD450-OD650 was determined.
The purity of recombinant hTGF-β2 was determined by SDS-PAGE of 6 µg reduced (+) and non-reduced (-) recombinant hTGF-β2 and staining overnight with Coomassie Blue.
Western blot analysis of extracts from HT-1080 cells untreated or treated with TGF-β2 for 15 minutes, using Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb #3108 (upper) and Smad2 (86F7) Rabbit mAb #3122 (lower).
With carrier: Lyophilized from a 0.22 μm filtered solution of 35% acetonitrile and 0.1% trifluoroacetic acid containing 20 μg BSA per 1 μg hTGF-β2. Carrier free: Lyophilized from a 0.22 μm filtered solution of 35% acetonitrile and 0.1% trifluoroacetic acid.
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 hTGF-β2. All lots are greater than 98% pure.
Recombinant hTGF-β2 contains no "tags" and the nonglycosylated protein has a calculated MW of 12,719. DTT-reduced protein migrates as a 12 kDa polypeptide and the non-reduced cystine-linked homodimer migrates as a 24 kDa protein. The expected amino-terminal ALDAA of recombinant hTGF-β2 was verified by amino acid sequencing.
The bioactivity of recombinant hTGF-β2 was was determined by assessing inhibition of IL-4 induced HT-2 cell proliferation. The ED50 of each lot is between 0.1-0.3 ng/ml.
Less than 0.01 ng endotoxin/1 μg hTGF-β2.
Recombinant human TGF-β2 (hTGF-β2) Ala303-Ser414 (Accession #NP_003229) was expressed in human 293 cells at Cell Signaling Technology.
TGF-β2 is produced by eosinophils, astrocytes, glioblastoma and other cancer derived cell types (1-6). TGF-β2 inhibits antigen-induced T-cell proliferation, inhibits epithelial cell proliferation, induces mesenchymal cell proliferation and has cell specific effects on apoptosis (1-8). TGF-β2 binds to TβRII and binding is promoted by TβRIII. TβRI then complexes with TβRII and TβRIII (3,5). Signaling involves phosphorylation of the SMAD proteins (2,3,9). TGF-β2 also activates Erk2, p38, and JNK pathways (9). Knockout of TGF-β2 in mice severely impacts heart, lung and eye development (10).
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