Analysis of extracellular matrix proteins produced by cultured cells
| العنوان: | Analysis of extracellular matrix proteins produced by cultured cells |
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| المؤلفون: | Turoverova Lv, Miralda I. Blinova, M.G. Khotin, Karl-Eric Magnusson, N. M. Yudintseva, D. Tentler, Pinaev Gp |
| المصدر: | Cell and Tissue Biology. 3:497-502 |
| بيانات النشر: | Pleiades Publishing Ltd, 2009. |
| سنة النشر: | 2009 |
| مصطلحات موضوعية: | biology, Cell, Cell Biology, Cell biology, Fibronectin, Extracellular matrix, chemistry.chemical_compound, Membrane, medicine.anatomical_structure, chemistry, Epidermoid carcinoma, Biochemistry, Cell culture, biology.protein, medicine, Sodium dodecyl sulfate, Polyacrylamide gel electrophoresis |
| الوصف: | The extracellular matrix (ECM) is a highly organized multimolecular structure essential for the vital functions of any organism. Although much of the data of extracellular matrix components has been accumulated, the isolation of an entire set of these proteins remains a complex procedure due to the high content of fibrillar proteins and proteoglycans, which form multidomain, netlike structures. In the study presented, we developed a method for isolating ECM proteins from cell cultures. Human epidermoid carcinoma cells A431 and fibroblasts obtained from normal and scar human skin were used. We showed that EDTA solution removed cells from culture plates without destroying the cell membranes. Subsequent treatment of remaining ECM proteins with acetic acid in order to dissociate collagen fibers significantly improved the fractioning of ECM proteins. The extraction of remaining proteins from the surface of the culture plate was preformed by a buffer developed based on Laemmli probe buffer. Using this method, we isolated ECM proteins synthesized by cultured cells, and the extracted proteins were suitable for future analysis by SDS PAGE and two-dimentional electrophoresis, as well as for identifying individual proteins by mass spectrometry. This study may allow us to compare assortments of ECM proteins isolated from different sources, and elucidate impact of various proteins on structure and property of extracellular matrix of investigated cells. |
| تدمد: | 1990-5203 1990-519X |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_________::ae30d9a83308d969e13575613b91a10b https://doi.org/10.1134/s1990519x09050137 |
| حقوق: | CLOSED |
| رقم الأكسشن: | edsair.doi...........ae30d9a83308d969e13575613b91a10b |
| قاعدة البيانات: | OpenAIRE |
Find this article in full text from Springer Verlag. | تدمد: | 19905203 1990519X |
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