Clinical Chemistry
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Clinical Chemistry 33: 2011-2018, 1987;
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Clinical Chemistry, Vol 33, 2011-2018, Copyright © 1987 by American Association for Clinical Chemistry

An examination of heart proteins by two-dimensional electrophoresis

E Gianazza, L Osio, G Grazioli, S Astrua-Testori, PG Righetti, R Accinni, I Renoldi and A Repossini
Faculty of Pharmacy, University of Milan, Italy.

We examined specimens from explanted human hearts by two-dimensional electrophoresis. The protocol selected includes: (a) solubilization of the sample in a urea-detergent mix; (b) charge fractionation in the presence of urea and nonionic detergent on a pH 4-10 immobilized pH gradient; (c) size fractionation on a polyacrylamide concentration gradient in the presence of sodium dodecyl sulfate; and (d) staining with silver nitrate. The method is sensitive enough for analysis of biopsies in the 1-3 mg range (wet tissue). We saw, for explanted hearts, variations in the protein pattern with the site of sample dissection. Results are presented for 11 explanted human hearts: one control organ and 10 pathological samples. The recorded pathologies included dilatative cardiomyopathy (six cases), valvulopathy (one case), ischemic cardiopathy (two cases), and graft rejection (one case). The patterns for whole extracts as well as for cytoplasmic proteins and myofibril components are compared. Extensive individual variability was observed both between control and pathological cases and among the abnormal samples.




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