Automatic transfer of polyacrylamide gels to protein chip plates for protein electroblotting

Akiko Okayama, Yoko Ino, Keita Mishima, Takeshi Okada, Y. Iwafune, Noriaki Arakawa, H. Kawasaki, H. Hirano
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Abstract

Protein chips are useful tools for profiling proteins and analyzing protein-protein interactions and post-translational modifications. In previous work, we developed a diamond-like carbon-coated stainless steel plate (DLC plate) as a novel protein chip plate. Gel-resolved proteins can be covalently immobilized on the surface of the DLC plate by electroblotting to produce a high-density protein chip. The proteins can then be identified by matrix-assisted laser desorption ionization/time-of-flight mass spectrometry (MALDI-TOF MS) using the plates. The interactions of the immobilized proteins with other proteins, and their post-translational modifications, can also be analyzed. However, to improve the efficiency and reproducibility of analyses using DLC plates, it is important to automate these analytical processes. Therefore, we developed a system for automatically transferring gels from the gel electrophoresis glass plates to the DLC plates for electroblotting. This is an essential first step toward complete automation of the production of high-density protein chips for immobilizing gel-resolved proteins.
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聚丙烯酰胺凝胶自动转移到蛋白质芯片板,用于蛋白质电印迹
蛋白质芯片是分析蛋白质、蛋白质相互作用和翻译后修饰的有用工具。在之前的工作中,我们开发了一种类金刚石碳涂层不锈钢板(DLC板)作为一种新型蛋白质芯片板。通过电印迹法将凝胶溶解蛋白共价固定在DLC板表面,制成高密度蛋白芯片。然后可以用基质辅助激光解吸电离/飞行时间质谱(MALDI-TOF MS)鉴定蛋白质。还可以分析固定蛋白与其他蛋白的相互作用及其翻译后修饰。然而,为了提高DLC板分析的效率和重现性,自动化这些分析过程是很重要的。因此,我们开发了一种系统,可以自动将凝胶从凝胶电泳玻璃板转移到DLC板上进行电印迹。这是实现用于固定化凝胶溶解蛋白的高密度蛋白芯片生产完全自动化的重要的第一步。
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