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

Akiko Okayama, Yoko Ino, Keita Mishima, Takeshi Okada, Y. Iwafune, Noriaki Arakawa, H. Kawasaki, H. Hirano
{"title":"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","authors":"Akiko Okayama, Yoko Ino, Keita Mishima, Takeshi Okada, Y. Iwafune, Noriaki Arakawa, H. Kawasaki, H. Hirano","doi":"10.2198/JELECTROPH.52.43","DOIUrl":null,"url":null,"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.","PeriodicalId":15059,"journal":{"name":"Journal of capillary electrophoresis","volume":"24 1","pages":"43-46"},"PeriodicalIF":0.0000,"publicationDate":"2008-06-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Automatic transfer of polyacrylamide gels to protein chip plates for protein electroblotting\",\"authors\":\"Akiko Okayama, Yoko Ino, Keita Mishima, Takeshi Okada, Y. Iwafune, Noriaki Arakawa, H. Kawasaki, H. Hirano\",\"doi\":\"10.2198/JELECTROPH.52.43\",\"DOIUrl\":null,\"url\":null,\"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.\",\"PeriodicalId\":15059,\"journal\":{\"name\":\"Journal of capillary electrophoresis\",\"volume\":\"24 1\",\"pages\":\"43-46\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2008-06-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of capillary electrophoresis\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.2198/JELECTROPH.52.43\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of capillary electrophoresis","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.2198/JELECTROPH.52.43","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

摘要

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。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Automatic transfer of polyacrylamide gels to protein chip plates for protein electroblotting
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.
求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
期刊最新文献
Electrophoretic extraction of protein complexes after separation and detection by a combined method of non-denaturing two-dimensional electrophoresis and reversible staining Use of Escherichia coli expression system for analyzing kinase motifs Proteomic analysis of spheroids of rhabdomyosarcoma cells cultured with decellularized muscle extracts Drug screening and kinase activity profiling of a novel patient-derived cell line of clear cell ovarian carcinoma Proteogenomic approach to drug targets in osteosarcomas with different original sites
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1