Fernandez J, Gharahdaghi F, Mische S M
The Rockefeller University Protein/DNA Technology Center, New York, NY 10021, USA.
Electrophoresis. 1998 May;19(6):1036-45. doi: 10.1002/elps.1150190619.
As the resource laboratory for Rockefeller University our emphasis continues to be on methodology development for the routine analysis of low abundance proteins isolated from native sources. In the past ten years, gel electrophoresis of proteins has become the method of choice for the preparation of microgram and submicrogram quantities of protein for primary structural characterization, and over 95% of the samples submitted for protein identification are either in a gel or bound to polyvinyl difluoride membranes (PVDF). As such, we employ multiple microanalytical approaches encompassing Edman sequence degradation, amino acid and matrix assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometric analysis to provide an integrated protein characterization of such samples. Here we describe the two major services we employ when providing protein identification from in-gel or PVDF-bound proteins.
作为洛克菲勒大学的资源实验室，我们继续将重点放在从天然来源分离的低丰度蛋白质常规分析的方法开发上。在过去十年中，蛋白质凝胶电泳已成为制备微克和亚微克量蛋白质以进行一级结构表征的首选方法，提交用于蛋白质鉴定的样本中超过95% 要么在凝胶中，要么与聚偏二氟乙烯膜（PVDF）结合。因此，我们采用多种微量分析方法，包括埃德曼序列降解、氨基酸分析以及基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱分析，以对这类样本进行综合蛋白质表征。在此，我们描述在从凝胶内或PVDF结合的蛋白质中进行蛋白质鉴定时所采用的两项主要服务。
