MOBILITY STUDY OF ALLYLAMINE BY FRAP

通过 FRAP 进行烯丙胺的迁移率研究

• 批准号: 7373162
• 负责人: STEVEN L. REGEN
• 金额: $ 0.34万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7373162
• 关键词: MOBILITY; STUDY; ALLYLAMINE; FRAP

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. We are interested to study the mobility of surface adsorbed polyallylamine using Fluorescence Recovery After Photobleaching (FRAP), a single molecule based technique. This technique gives us a direct measure of the rates of lateral transport of proteins and lipids in cell membranes. For us, the three issues of greatest interest are: (1) Does poly(allylamine) come off of a layer-by-layer surface [with an underlayer of poly(styrenesulfonate)] under any conditions; e.g., acid, base high salt concentration, (2) is there any lateral mobility of the outer poly(allyamine) layer under any conditions; i.e., FRAP within this outer layer, and (3) how does our novel covalently-cross-linked poly(allylamine) surface compare with conventional ionically-cross-linked, layer-by-layer surfaces in terms of stability and lateral mobility. For this the polyallylamine is labeled with a fluorescent dye that can be excited by the 514 nm line of an Argon-ion laser which in turn is coupled to a confocal microscope.

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。我们感兴趣的是利用光漂白后荧光恢复（FRAP），一种基于单分子的技术来研究表面吸附的聚烯丙胺的迁移率。这项技术使我们能够直接测量细胞膜中蛋白质和脂质的横向运输速率。对我们来说，最感兴趣的三个问题是：(1)在任何条件下，聚（烯丙胺）是否从一层一层的表面（下层是聚（苯乙烯磺酸盐））脱落；例如，酸、碱高盐浓度，(2)在任何条件下，外层聚丙烯胺层是否存在任何横向迁移性；(3)在稳定性和横向迁移率方面，我们的新型共价交联聚（烯丙胺）表面与传统的离子交联层表面相比如何。为此，用荧光染料标记聚烯丙胺，荧光染料可以被氩离子激光的514 nm线激发，而氩离子激光又与共聚焦显微镜耦合。