Suppr超能文献

优化垂直 mini-凝胶电泳系统中的血管性血友病因子多聚体分析:一种快速方法。

Optimization of von Willebrand factor multimer analysis in vertical mini-gel electrophoresis systems: A rapid procedure.

机构信息

Instituto Butantan, Laboratório de Fisiopatologia, São Paulo, SP, Brazil; Faculdade de Medicina, Universidade de São Paulo, São Paulo, SP, Brazil.

Fundação Pró-Sangue, Hemocentro de São Paulo, Faculdade de Medicina da Universidade de São Paulo, São Paulo, SP, Brazil.

出版信息

Thromb Res. 2019 Mar;175:76-83. doi: 10.1016/j.thromres.2019.01.018. Epub 2019 Jan 30.

Abstract

Von Willebrand disease (VWD) is a common cause of bleeding worldwide. Analysis of von Willebrand factor (VWF) multimer distribution (VWF:MD) is essential to properly classify and treat different types of VWD, and it is performed using a SDS agarose gel electrophoresis followed by Western blotting, a handmade technique that demands days to be completed and requires skillful execution. Aiming both to facilitate gel production and to shorten the preparation time, we developed an uncomplicated technique to provide agility in the analysis of VWF:MD, so that it can be easily accomplished in the routine practice of hemostasis laboratories. On that account, we used a commercial vertical mini-gel electrophoresis system for SDS-PAGE and a semi-dry transfer system, which allowed us to analyze VWF:MD of various samples in a period shorter than 12 h. This technique differentiated VWF:MD in human and animal plasmas under normal, congenital and acquired (experimental envenomation by Bothrops jararaca snake) conditions. This optimized method is cheap, rapid, reproducible, easy to be performed, and uses electrophoresis and Western blotting systems available in most laboratories. All these advantages encourage hemostasis professionals to use it in their routine practices. In order to facilitate the setup and accomplishment of the whole procedure step by step, videos were appended to the article.

摘要

血管性血友病(von Willebrand disease,VWD)是一种在全球范围内较为常见的出血性疾病。分析血管性血友病因子(von Willebrand factor,VWF)多聚体分布(VWF:MD)对于正确分类和治疗不同类型的 VWD 至关重要,通常采用十二烷基硫酸钠琼脂糖凝胶电泳(SDS agarose gel electrophoresis)结合免疫印迹法(Western blotting)进行分析,这是一种手工操作技术,需要数天才能完成,且对技术要求较高。为了简化凝胶制备过程并缩短准备时间,我们开发了一种简便的技术,以提高 VWF:MD 分析的灵活性,使其能够在止血实验室的常规实践中轻松完成。为此,我们使用商业的垂直迷你电泳系统进行 SDS-PAGE 电泳和半干式转印系统,使我们能够在 12 小时内分析各种样本的 VWF:MD。该技术可区分正常、先天性和获得性(巴西矛头蝮蛇毒液诱导的实验性中毒)条件下人类和动物血浆中的 VWF:MD。这种优化方法成本低、速度快、重现性好、易于操作,并且使用了大多数实验室都具备的电泳和 Western blotting 系统。所有这些优势都鼓励止血专业人员在日常实践中使用它。为了方便逐步设置和完成整个操作过程,我们在文章中附加了视频。

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验