Miniaturized two-dimensional gel electrophoresis of high-molecular-weight proteins using low-concentration multifilament-supporting gel for isoelectric focusing
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概要
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A miniaturized two-dimensional gel electrophoresis (2-DE) system was constructed with a multifilament-supporting (MFS) gel as a first-dimensional gel for isoelectric focusing (IEF). The MFS gel was 1 mm in diameter, had a multifilament yarn of acrylic as a gel-support, and could provide low-concentration (2.5%T) polyacrylamide with sufficient mechanical strength, resulting in fast IEF, and clear 2-D spots due to the relatively large pore and improved transfer efficiency from the first-dimensional gel to the second-dimensional one of especially high-molecular-weight (HMW) proteins. In this study, by using the MFS gel of 4 cm in length as a miniaturized IEF gel, three colored proteins were separated in 8 min, which was about twice faster than using the MFS gel of standard length (7 cm). The second-dimensional gel was also miniaturized to be 55 mm wide, 40 mm long and 0.75 mm thick. The low-concentration MFS gel after IEF was easily transferred onto the top of the miniaturized second-dimensional gel with a tweezers. By using the miniaturized 2-DE system, the HMW size marker was separated in 30 min by IEF and in 25 min by native-polyacrylamide gel electrophoresis (Native-PAGE), which was at least twice faster than by using the standard size 2-DE system using MFS gel. This miniaturized 2-DE system could be expected as a useful separation method for fast protein diagnosis and screening.
- 日本電気泳動学会の論文
- 2009-12-01
著者
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角田 欣一
群馬大学大学院工学研究科応用化学・生物化学専攻
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HOTTA Hiroki
Department of Chemistry and Chemical Biology, Gunma University
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TSUNODA Kin-ichi
Department of Chemistry and Chemical Biology, Gunma University
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LI Jinxiang
Department of Chemistry, Liaoning Normal University
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Hotta Hiroki
Department Of Chemistry And Chemical Biology Gunma University
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小竹 玉緒
東大院・工
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Takiguchi Hiromi
School Of Engineering The University Of Tokyo
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Li Jinxiang
Department Of Chemistry Liaoning Normal University
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Kojima Yasuaki
Mitachi Co. Ltd.
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Hotta Hiroki
Department Of Biochemistry Kyoto Prefectural University
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ODAKE Tamao
School of Engineering, The University of Tokyo
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SAHEKI Toshihiko
Department of Chemistry and Chemical Biology, Gunma University
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KAMEZAWA Kazutoshi
Department of Chemistry and Chemical Biology, Gunma University
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MIYATA Kozue
Department of Chemistry and Chemical Biology, Gunma University
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ONDA Koki
Textile Research Institute of Gunma
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Miyata Kozue
Department Of Chemistry And Chemical Biology Gunma University
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Tsunoda Kin-ichi
Department Of Chemistry And Chemical Biology Gunma University
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Tsunoda Kin-ichi
Department Of Applied Chemistry Faculty Of Engineering Gunma University
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Saheki Toshihiko
Department Of Chemistry And Chemical Biology Gunma University
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Kamezawa Kazutoshi
Department Of Chemistry And Chemical Biology Gunma University
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SAHEKI Toshihiko
Department of Biochemistry, Faculty of Medicine, Kagoshima University
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Hotta Hiroki
Department of Biochemistry and Engineering, Faculty of Engineering, Tohoku University
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