优化用于通过 LC-MS/MS 分析 HeLa S3 细胞的细胞裂解和蛋白质消化方案。

Optimization of cell lysis and protein digestion protocols for the analysis of HeLa S3 cells by LC-MS/MS.

机构信息

Department of Pathology, Children's Hospital Boston and Harvard Medical School, Boston, MA 02115, USA.

出版信息

Proteomics. 2011 Dec;11(24):4726-30. doi: 10.1002/pmic.201100162. Epub 2011 Nov 23.

DOI:10.1002/pmic.201100162

PMID:22002805

Abstract

In order to maximize the number of proteins identified from Hela S3 cell lysate we tested various cell lysis, protein precipitation and digestion protocols. First, we compared three different lysis buffers, two mechanical cell disruption methods and two precipitation methods. Then, we tested six different in-solution digestion protocols, three different in-gel digestion protocols and ten different peptide extraction protocols. The result is a proposal for an optimized protocol to prepare the whole cell lysate samples from HeLa S3 cells.

摘要

为了从 Hela S3 细胞裂解物中鉴定出尽可能多的蛋白质，我们测试了各种细胞裂解、蛋白质沉淀和消化方案。首先，我们比较了三种不同的裂解缓冲液、两种机械细胞破碎方法和两种沉淀方法。然后，我们测试了六种不同的溶液内消化方案、三种不同的胶内消化方案和十种不同的肽提取方案。最终提出了一个优化方案，用于从 Hela S3 细胞制备全细胞裂解物样品。