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Influence of gel dimensions on resolution and sample throughput on two-dimensional gels

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Abstract

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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Abbreviations

2-D PAGE:

two-dimensional polyacrylamide gel electrophoresis

IEF:

isoelectrofocusing

IPG:

immobilized pH gradient

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Authors and Affiliations

  1. Department of Biotechnology, SongDo TechnoPark, 7-50 Songdo-Dong, Yeonsu-Gu, Incheon, Korea, 406-840

    KiBeom Lee

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

    KyungBae Pi

  3. School of Agricultural Biotechnology, Seoul National University, Seoul, 151-742, Korea

    Hong-Gu Lee

Authors
  1. KiBeom Lee
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  2. KyungBae Pi
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  3. Hong-Gu Lee
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Corresponding authors

Correspondence to KiBeom Lee or Hong-Gu Lee.

Additional information

Published in Russian in Biokhimiya, 2008, Vol. 73, No. 4, pp. 524–529.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM07-291, January 27, 2008.

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Lee, K., Pi, K. & Lee, HG. Influence of gel dimensions on resolution and sample throughput on two-dimensional gels. Biochemistry Moscow 73, 427–431 (2008). https://doi.org/10.1134/S000629790804007X

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  • DOI: https://doi.org/10.1134/S000629790804007X

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