Application of two‐dimensional electrophoresis for plasma of normal and diabetic rats

• Published in: The FASEB journal 2013-04, Vol.27 (S1)
• Main Authors: Braga, Camila Pereira, Pozzi, Carla Mariane Costa, Vieira, José Cavalcante Souza, do Carmo Federici Padilha, Cilene, Okoshi, Marina Politi, Fernandes, Ana Angélica Henrique, de Magalhães Padilha, Pedro
• Format: Article
• Language: English
• ISSN: 0892-6638; 1530-6860
• DOI: 10.1096/fasebj.27.1_supplement.lb137

Abstract only The aim of this study was to compare plasma gels of normal and diabetic rats by two‐dimensional electrophoresis. Experimental animals were randomly divided into two groups (n = 8): G1 = control and G2 = diabetic rats (induced by intraperitoneal administration of streptozotocin in a single dose concentration of 60 mg/kg body weight). A pool of plasma samples were diluted in a specific buffer and applied to IEF (isoelectric focusing) strips with pH in the range of 4–7. Once hydrated for 12h, the strips were brought to the IEF system for running the first dimension. After this step, the strips were equilibrated in a specific buffer for 15 minutes and applied onto polyacrylamide gels at 10% w/v for running the second dimension. At the end of the separation steps (performed in duplicate), the protein spots were fixed in the gel and then stained with Coomassie Blue. The gels were scanned and analyzed using an image processor. For the image analysis, it was found that the protein spots for both experimental groups were more concentrated in the range of 24–76 kDa; 416 spots were found in G1 and 513 in G2, with a percentual matching of 53 and 43% for G1 and G2 respectively. These matching values show that there was some difference between the treatments. There is great interest in the plasma proteome as a discovery source of new biomarkers in diabetes as it is thought to contain subsets of other tissue proteomes. Financial Support: FAPESP 2011/21672–1