Molecular Micellar-Polyacrylamide Gel Electrophoresis (MoMi-PAGE)

分子胶束-聚丙烯酰胺凝胶电泳 (MoMi-PAGE)

• 批准号: 7347388
• 负责人: ISIAH M WARNER
• 金额: $ 17.44万
• 依托单位: LOUISIANA STATE UNIV A&M COL BATON ROUGE
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-15 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7347388
• 关键词: Acrylamide; Acrylamides; Affect; Architecture; Area; Arterial Fatty Streak; Arteries; Belief; Biological; Blood; Blood capillaries; Charge; Chemicals; Chromatography; Class; Complex Mixtures; Data; Diagnosis; Disease; Early Diagnosis; Experimental Designs; Fluorescent Probes; Funding; Goals; Grant; Health; Heart; Human; Hydrophobicity; Individual; Laboratories; Lead; Micelles; Molecular; Polyacrylamide Gel Electrophoresis; Polymers; Procedures; Proteins; Reagent; Research; Sampling; Sodium Dodecyl Sulfate; Solutions; Solvents; Structure; System; Techniques; Technology; Time; Tubular formation; United States National Institutes of Health; base; capillary; chiral molecule; concept; design; desire; gel electrophoresis; human disease; improved; interest; molecular recognition; novel; stem; success; surfactant

DESCRIPTION (provided by applicant): This application outlines research which focuses on the use of molecular micelles to replace sodium dodecyl sulfate (SDS) and other conventional micelles in poly- acrylamide gel electrophoresis. The overall hypothesis of this research stems from our belief that molecular micelles should provide better PAGE separation of proteins than SDS or other conventional micelles. This hypothesis is based upon previous separation strategies developed in our laboratory for separation of chiral molecules. In addition, we have preliminary data provided in this application which supports this hypothesis. Thus, the overall objective of the research described here is to examine better separation strategies for analyses of protein samples using molecular micellar-polyacrylamide gel electrophoresis (MoMi-PAGE). To achieve this objective, this application is divided into three specific aims. The first specific aim is to understand the parameters which contribute to PAGE separations using molecular micelles as alternative reagents to SDS and other conventional micelles which are frequently used in PAGE separations. This first specific aim builds upon expertise developed over the past several years in our laboratory. Under this specific aim, we will explore various strategies with molecular micelles of specifically designed architectures to optimize separations for different classes of proteins using PAGE separations. Our second specific aim is to understand the mechanisms of our separations at the molecular level since previous success with our separation research has been the result of treating our separation problems as a molecular recognition problem. To achieve this aim, we will use fluorescent probes and experimental design to understand these parameters which contribute to optimal separations. Our third specific aim is to apply the technology that we have developed to the analyses of proteins associated with the formation of atherosclerotic plaques and proteins associated with other biological systems of interest. This latter specific aim is mostly descriptive of the kinds of application that we will ultimately explore since it is unlikely that we would get to such applications during the proposed two-year time frame of this application. Finally, it is noted that our previous studies under NIH funding, as well as the individual expertise of the collaborators brought together for this research, enhances the chance for success of the overall goals outlined in this application. Protein separations are very important to understanding many diseases which affect human health. Therefore, it is important to develop better separation and identification procedures for analyses of proteins. The most widely used protein separation technique is called sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE) or (SDS-PAGE), which employs SDS for solubilization and separation of proteins on an electric grid. We propose to develop new reagents, call molecular micelles, to replace SDS and other surfactants in order to improve PAGE separation of proteins.

描述（由申请人提供）：该申请概述了研究的研究，该研究侧重于使用分子胶束替代硫酸钠（SDS）（SDS）和其他常规胶束中的聚丙烯酰胺凝胶电泳。这项研究的总体假设源于我们相信分子胶束应比SDS或其他常规胶束提供更好的蛋白质分离。该假设是基于我们实验室中先前针对手性分子分离的分离策略。此外，我们在本应用程序中提供了支持该假设的初步数据。因此，此处描述的研究的总体目标是检查使用分子胶束 - 聚丙烯酰胺凝胶电泳（MOMI-PAGE）分析蛋白质样品的更好的分离策略。为了实现这一目标，该应用程序分为三个特定目标。第一个具体目的是了解使用分子胶束作为SDS和其他常规胶束的替代试剂，这些参数有助于页面分离，这些试剂经常用于页面分离。这个第一个特定的目标是基于在过去几年中在我们的实验室中发展的专业知识。在这个具体目标下，我们将使用专门设计的架构进行分子胶束探索各种策略，以使用页面分离来优化不同类别的蛋白质的分离。我们的第二个具体目的是了解分子水平分离的机制，因为我们先前的分离研究成功一直是将分离问题视为分子识别问题的结果。为了实现这一目标，我们将使用荧光探针和实验设计来了解这些参数，从而有助于最佳分离。我们的第三个具体目的是将我们开发的技术应用于与与其他感兴趣的生物学系统相关的动脉粥样硬化斑块和蛋白质形成相关的蛋白质分析。后一个特定的目的主要是描述我们最终将探索的应用程序的描述，因为我们不太可能在本申请的两年时间内获得此类申请。最后，人们指出的是，我们以前根据NIH资助的研究以及合作者的个人专业知识汇集了这项研究，这增加了该应用程序中概述的整体目标的成功机会。 蛋白质分离对于理解影响人类健康的许多疾病非常重要。因此，为蛋白质分析制定更好的分离和识别程序很重要。使用最广泛的蛋白质分离技术称为十二烷基硫酸钠（SDS）聚丙烯酰胺凝胶电泳（PAGE）或（SDS-PAGE），该胶质凝胶电泳（PAGE）或（SDS-PAGE）使用SDS来溶解和在电网上的蛋白质分离。我们建议开发新的试剂，称分子胶束，以取代SDS和其他表面活性剂，以改善蛋白质的页面分离。