Abstract
Radioiodinated transforming growth factors ( TGF-βs) are critical reagents for functional studies on TGF-β ligand-receptor binding (1–5), TGF-β-binding protein interactions (6–9), ligand uptake, degradation and clearance (1,10,11), titering of TGF-β neutralizing antibodies (9,10), and for quantitation of TGF-βs by radioimmuno and radioreceptor assays (12,13). Several methods have been developed for the iodination of TGF-βs, namely chloramine T, lac-toperoxidase, and Bolton-Hunter (1,3,4,14). Although the latter method is gentler and believed to yield radiolabeled TGF-βs with greater biological activity than the first two, it is much more expensive, labor intensive and yields a poorly labeled product. Chloramine T is currently the most common method for iodination of TGF-βs, most likely because of its great efficiency of labeling. However, iodination with chloramine T can cause oxidative degradation of TGF-βs with excess treatment. For this reason, successful iodination of TGF-βs requires strict adherence to the amount of chloramine T used and the duration of treatment. Frolik et al. (1) have shown that three consecutive short treatments with low levels of chloramine T gives the best labeling with little oxidation of TGF-β. With the chloramine T method outlined later, all three isoforms of TGF-β can be iodinated with near-equal efficiency.
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References
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Danielpour, D. (2000). Methods for Iodination of Active TGFβ, TGFβ Receptor Crosslinking and Immunoprecipitation of TGFβ-Receptor Complexes. In: Howe, P.H. (eds) Transforming Growth Factor-Beta Protocols. Methods in Molecular Biology™, vol 142. Humana Press. https://doi.org/10.1385/1-59259-053-5:29
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DOI: https://doi.org/10.1385/1-59259-053-5:29
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