一种用于毫秒时间范围内光亲和标记研究的停流装置。在烟碱型乙酰胆碱受体研究中的应用。
A stopped-flow apparatus for photoaffinity labeling studies in the milliseconds time range. Application in investigations of the nicotinic acetylcholine receptor.
作者信息
Fahr A, Hucho F
出版信息
J Neurosci Methods. 1986 Mar;16(1):29-38. doi: 10.1016/0165-0270(86)90005-1.
A photoaffinity labeling method is described to label a protein covalently in various transient covalent states. The method uses a combination of an especially adapted stopped-flow apparatus with a Q-switched Nd: YAG DCR-2A laser (wavelength 266 nm, i.e. four-fold the primary frequency, pulse duration 4 ns, pulse energy 15 mJ). The construction of the mixing cell, the triggering device and the set-up for determining the dead time of the stopped-flow apparatus is described. The dead time is 2.4 ms. In combination with a specific photolabel the method has been used for labeling functional states (resting, activated, desensitized, antagonist-blocked) of the nicotinic acetylcholine receptor from Torpedo marmorata electric tissue.
本文描述了一种光亲和标记方法，用于在各种瞬态共价状态下对蛋白质进行共价标记。该方法将特别改装的停流装置与调Q钕：钇铝石榴石DCR - 2A激光（波长266 nm，即基频的四倍，脉冲持续时间4 ns，脉冲能量15 mJ）结合使用。文中描述了混合池的构造、触发装置以及用于确定停流装置死时间的设置。死时间为2.4 ms。结合特定的光标记物，该方法已用于标记来自电鳐电组织的烟碱型乙酰胆碱受体的功能状态（静息、激活、脱敏、拮抗剂阻断）。
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