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Improved method for immunostaining of mucin separated by supported molecular matrix electrophoresis by optimizing the matrix composition and fixation procedure†
Article first published online: 24 JUN 2011
Copyright © 2011 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Volume 32, Issue 14, pages 1829–1836, July 2011
How to Cite
Matsuno, Y.-K., Dong, W., Yokoyama, S., Yonezawa, S., Saito, T., Gotoh, M., Narimatsu, H. and Kameyama, A. (2011), Improved method for immunostaining of mucin separated by supported molecular matrix electrophoresis by optimizing the matrix composition and fixation procedure. ELECTROPHORESIS, 32: 1829–1836. doi: 10.1002/elps.201000608
- Issue published online: 18 JUL 2011
- Article first published online: 24 JUN 2011
- Manuscript Accepted: 23 MAR 2011
- Manuscript Revised: 17 MAR 2011
- Manuscript Received: 11 NOV 2010
- New Energy and Industrial Technology Development Organization (NEDO)
- O-linked glycans;
Mucins are a family of heavily glycosylated high molecular mass proteins that have great potential as novel clinical biomarkers for the diagnosis of various malignant tumors. Supported molecular matrix electrophoresis (SMME) is a new type of membrane electrophoresis that can be used to characterize mucins. In SMME, mucins migrate in a molecular matrix supported by membrane materials. Here, we have developed an immunostaining method for the identification of SMME-separated mucins. The novel method involves stably fixing the mucins onto the SMME membrane and optimizing the molecular matrix for the fixation process. We applied this technique for the detection of MUC1 produced from three cancer cell lines (T47D, HPAF-II and BxPC3) and also analyzed their O-linked glycans by mass spectrometry. Our results revealed that properties of the MUC1 molecules from the three cell lines are different in terms of migrating position in SMME and glycan profile. The present method allows simple and rapid characterization of mucins in terms of both glycans and core proteins. The method will be a useful tool for the exploration of mucin alterations associated with various diseases such as cancer.