Influence of Gel Dimensions on Resolution and Sample Throughput on Two-Dimensional Gels

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KiBeom Lee^1*, KyungBae Pi², and Hong-Gu Lee^3*

¹Department of Biotechnology, SongDo TechnoPark, 7-50 Songdo-Dong, Yeonsu-Gu, Incheon, Korea 406-840; fax: 82-32-260-0769; E-mail: klee02@empal.com

²Research Institute, AdipoGen, Korea University, College of Life Science and Biotechnology, Rm#538, 5-Ka, Anam-Dong, Sungbuk-Gu, Seoul, Korea 136-701

³School of Agricultural Biotechnology, Seoul National University, Seoul 151-742, Korea; fax: 82-2-875-7340; E-mail: hglee66@hotmail.com

^* To whom correspondence should be addressed.

Received September 3, 2007; Revision received September 27, 2007

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To achieve high throughput and economical format of 2-D PAGE, comparison between gel size and resolution was conducted on human breast carcinoma cell line (MCF-7/AZ) proteins. SDS gel length showed a weaker influence of separation length on resolution in the second dimension, and there was little benefit of separation distances greater than 15 to 19 cm. IPG strip separation distances were very important with dramatic increase in resolution of longer gels compared with smaller gels, and maximal resolution was obtained using 18- and 24-cm IPG strips. Loading optimal amount of proteins on 2-D gels can also increase the number of detected spots. Therefore, taken together, compromise 2-D gels are crucial for higher capacity and higher throughput.

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KEY WORDS: two-dimensional polyacrylamide gel electrophoresis, immobilized pH gradient (IPG) strip length, SDS gel length/high throughput, resolution

DOI: 10.1134/S000629790804007X

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