TECHNICAL ADVANCE: Free-flow electrophoresis for purification of plant mitochondria by surface charge
作者： Holger Eubel , Chun Pong Lee , John Kuo , Etienne H Meyer , Nicolas L Taylor
DOI: 10.1111/J.1365-313X.2007.03253.X
关键词: Biochemistry 、 Centrifugation 、 Proteomics 、 Free-flow electrophoresis 、 Gel electrophoresis 、 Biology 、 Peroxisome 、 Proteome 、 Electrophoresis 、 Surface charge 、 Molecular biology
摘要: Sample purity is the key for a successful in-depth analysis of any given subcellular proteome. The suitability free-flow electrophoresis to assist conventional, centrifugation-based techniques in preparation plant mitochondria from green and non-green tissue was assessed by various means, including functional assays, immunoblots, electron microscopy differential gel electrophoresis. Results indicated significant increase mitochondrial samples, highlighted specific contaminants previously reported as proteins, also pointed new means separating plastids peroxisomes organellar extracts exploiting differences surface charge. This approach has potential allow deeper more comprehensive investigation Arabidopsis proteomes, providing second dimension separation based on charge addition conventional centrifugation purification protocols relying size density.
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