Extraction and Separation Analysis of Total Protein from the Bark of Apple Branches
? These authors contributed equally to this paper
Received date: 2014-10-27
Accepted date: 2015-01-27
Online published: 2015-09-06
We aimed to establish a suitable extraction method for analyzing total protein in the bark of apple branches. The branches of an 8-year-old cultivar (Huayue) were used as material. We compared 3 different extraction methods, including TCA-acetone precipitation (A), methanol/ammonium acetate precipitation (B), and improved Tris-phenol extraction (C). We optimized the extraction procedures and established an ideal method for analyzing the total protein in the bark of apple branches. The improved Tris-phenol extraction method could reveal a good map of 993 protein spots, significantly more than the other two methods: 418 and 674 protein spots were obtained with TCA-acetone precipitation and methanol/ammonium acetate precipitation, respectively. Furthermore, the gel background was clear, and the protein spots were better focused than with the two other methods. After comparing sample loading quantity, we selected 800 μg as the ideal loading quantity for our study. Furthermore, we selected several proteins for MALDI-TOF-TOF/MS analysis and database searching, and the protein spots were finally identified. In this study, we optimized the extraction procedures and sample loading quantity for total protein and finally obtained an ideal 2-D electrophoresis map. Our study may provide a good basis for further proteome analysis of apple branches.
Caixia Zhang, Yi Tian, Liyi Zhang, Long Xiao, Peihua Cong . Extraction and Separation Analysis of Total Protein from the Bark of Apple Branches[J]. Chinese Bulletin of Botany, 2015 , 50(6) : 739 -745 . DOI: 10.11983/CBB14186
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