Western blot analysis of extracts from serum-starved HeLa cells treated for 30 minutes with various concentrations of Human TGF-β3 using Phospho-Smad2 (Ser465/467) (138D4) Rabbit mAb #3108 (upper) and Smad2 (86F7) Rabbit mAb #3122 (lower).Learn more about how we get our images
Recombinant human TGF-β3 expressed in E. coli
Recombinant human TGF-β3 expressed in E. coli and is supplied in a lyophilized form. A greater than 95% purity was determined by reverse-phase HPLC and SDS-PAGE.
Specific activity was determined to be less than 0.05 ng/ml.
The working concentration of TGF-β3 generally ranges from 10-50 ng/ml.
Lyophilized product is very stable at -20°C. It is recommended to reconstitute with sterile water at a concentration of 0.1 mg/ml, which can be further diluted in aqueous solutions as needed. Addition of a carrier protein (0.1% HSA or BSA) is recommended for long-term storage.
Transforming growth factor-β (TGF-β) superfamily members are critical regulators of cell proliferation and differentiation, developmental patterning and morphogenesis, and disease pathogenesis (1-4). TGF-β elicits signaling through three cell surface receptors: type I (RI), type II (RII), and type III (RIII). Type I and type II receptors are serine/threonine kinases that form a heteromeric complex. In response to ligand binding, the type II receptors form a stable complex with the type I receptors allowing phosphorylation and activation of type I receptor kinases (5). The type III receptor, also known as betaglycan, is a transmembrane proteoglycan with a large extracellular domain that binds TGF-β with high affinity but lacks a cytoplasmic signaling domain (6,7). Expression of the type III receptor can regulate TGF-β signaling through presentation of the ligand to the signaling complex. The only known direct TGF-β signaling effectors are the Smad family proteins, which transduce signals from the cell surface directly to the nucleus to regulate target gene transcription (8,9).
Three isoforms of TGF-β, designated TGF-β1, TGF-β2 and TGF-β3, are encoded by distinct genes and are expressed in a tissue specific manner (10). Each isoform is synthesized as a larger precursor protein containing a propeptide region that is removed prior to secretion. Mature TGF-β contains two polypeptides linked by disulfide bonds to form a protein of approximately 25 kDa.
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