Articles
A 2D-PAGE ANALYSIS OF PROTEIN DEPOSITION DURING THE DEVELOPMENT OF PHYLLOCLADS OF OPUNTIA FICUS-INDICA
Article number
728_14
Pages
111 – 116
Language
English
Abstract
A method for the extraction and analysis of proteins by two-dimensional electrophoresis from phylloclads of the cactus pear was developed.
This method is based on the extraction of proteins from freeze-dried phylloclads in 50 mM pyridine, 10 mM thiourea, 1% SDS, pH 5.0, and precipitation of the extracted proteins with 10% TCA in acetone.
Proteins were separated in the first dimension by isoelectric focusing in Immobiline DryStrips, pH 3-10. The second dimension of electrophoresis was performed in a uniform 15% separating gel at 25ºC and a under constant current of 25 mA. The efficiency of this protocol was demonstrated by comparison of the cactus pear proteome at four developmental stages of the phylloclads.
This method is based on the extraction of proteins from freeze-dried phylloclads in 50 mM pyridine, 10 mM thiourea, 1% SDS, pH 5.0, and precipitation of the extracted proteins with 10% TCA in acetone.
Proteins were separated in the first dimension by isoelectric focusing in Immobiline DryStrips, pH 3-10. The second dimension of electrophoresis was performed in a uniform 15% separating gel at 25ºC and a under constant current of 25 mA. The efficiency of this protocol was demonstrated by comparison of the cactus pear proteome at four developmental stages of the phylloclads.
Authors
E.A..R Vasconcelo, P.A.S. Souza, R.M. Liamoca Zarate, F.A.P. Campos
Keywords
cactus pear, forage crops, bi-dimensional electrophoresis, protein deposition, Cactaceae
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