[40] In vivo radiolabeling of human IFN-β
| Autor: | Diana Fahey, E. Knight |
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| Rok vydání: | 1986 |
| Předmět: | |
| DOI: | 10.1016/0076-6879(86)19042-2 |
| Popis: | Publisher Summary This chapter discusses the in vivo radiolabeling of human interferon β (IFN-β). Human IFN-β can be labeled with [35S]methionine in vivo to a specific radioactivity that is useful for many experiments. The advantage of in vivo radiolabeling is that all labeled molecules should have the same specific biological activity as the unlabeled molecules. Human diploid fibroblast cells are grown in 60-mm plastic culture dishes. IFN-β is assayed by a microtiter method with human diploid fibroblast cells and vesicular stomatitis virus as the challenge virus. The 35S-labeled IFN-β is purified to near homogeneity by chromatography on Blue Sepharose. The in vivo labeling of IFN-β with 35S-labeled methionine, and subsequent purification yields 35S-labeled IFN-β that can be used in various biochemical studies. 35S-labeled IFN-β has been used to investigate the removal of carbohydrate by glycosidoses from IFN-β and to study the biosynthesis of IFN-β in human diploid fibroblasts. |
| Databáze: | OpenAIRE |
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