On-Chip Identification and Interaction Analysis of Gel-Resolved Proteins Using a Diamond-like Carbon-Coated Plate

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• 作者：Yuko Iwafune ; Jian-Zhong Tan ; Yoko Ino ; Akiko Okayama ; Yuji Ishigaki ; Koji Saito ; Nobutake Suzuki ; Mikiko Arima ; Mitsuyoshi Oba ; Shuichi Kamei ; Michifumi Tanga ; Takeshi Okada ; Hisashi Hirano
• 刊名：Journal of Proteome Research
• 出版年：2007
• 出版时间：June 2007
• 年：2007
• 卷：6
• 期：6
• 页码：2315 - 2322
• 全文大小：593K
• 年卷期：v.6,no.6(June 2007)
• ISSN：1535-3907

文摘

We developed a novel protein chip made of a diamond-like, carbon-coated stainless steel plate (DLCplate), the surface of which is chemically modified with N-hydroxysuccinimide ester. To produce ahigh-density protein chip using the DLC plate, proteins separated by SDS gel electrophoresis or two-dimensional electrophoresis were electroblotted onto the DLC plate and immobilized covalently. A highblotting efficiency (25-70%) for transferring proteins from the gels onto the DLC plates was achievedby improvement of the electrophoresis device and electroblotting techniques. With the use of the DLCplate, we developed novel techniques to identify proteins immobilized on the chip and to detect protein-protein interactions on the chip by mass spectrometric analysis. We also developed a technique toidentify post-translationally modified proteins, such as glycoproteins, on the protein chip.