Login
欢迎浏览恩派尔生物资料网
我要投稿 请登录 免费注册 安全退出

您现在的位置是: 首页 > 实验方法 > 细胞技术

细胞技术

Liquid Chromatography MALDI MS/MS for Membrane Proteome Analysis

2025-03-07 细胞技术 加入收藏
Liquid chromatography (LC) can be combined with matrix-assisted laser-desorption

Liquid chromatography (LC) can be combined with matrix-assisted laser-desorption ionization (MALDI) mass-spectrometry (MS) by using automated off-line fraction collection of eluates onto a MALDI plate. Commercial tandem MS systems are available for generating product ion spectra of MALDI-produced peptide ions. The MALDI MS/MS spectra can be searched against a proteome database for protein identification. In this chapter, a protocol of sequential solubilization and digestion of membrane proteins involving methanol- and SDS-assisted trypsin digestion and microwave-assisted acid hydrolysis is presented. The process of LC eluate deposition onto a MALDI plate along with practical considerations for achieving optimal performance of eluate deposition is described. Issues related to MALDI MS and MS/MS spectral acquisition are discussed. Database searching and manual inspection of MS/MS spectra of singly charged MALDI-produced peptide ions for positive protein identification are also addressed.

文章底部广告位

文章评论

加载中~