Journal of Proteomics & Bioinformatics
Open Access
ISSN: 0974-276X
ISSN: 0974-276X
Edison T. Liu
Singapore
Research Article
A Chloroform-Assisted Protein Isolation Method Followed by Capillary NanoLC-MS Identify Estrogen-Regulated Proteins from MCF7 Cells
Author(s): Adaikkalam Vellaichamy, Chin-Yo Lin, Thin Thin Aye, Govindarajan R. Kunde, Alexey I. Nesvizhskii, Edison T. Liu and Siu Kwan SzeAdaikkalam Vellaichamy, Chin-Yo Lin, Thin Thin Aye, Govindarajan R. Kunde, Alexey I. Nesvizhskii, Edison T. Liu and Siu Kwan Sze
Most commonly reported non-commercial protein isolation methods require the use of detergents and/or chaotropes for better yield, and they all need additional Purification or clean-up steps for subsequent mass spectrometry analysis. Moreover, there is no simple procedure available for obtaining both soluble and membrane proteins from the same sample. Here we describe a simple and detergent-free chloroform-assisted protein isolation (ChlAPI) method for mammalian cells and tissues, and demonstrated its suitability to mass spectrometry based proteome analysis. In this single-step method, cultured cells or grounded tissue were mixed in 10% chloroform in ammonium bicarbonate buffer to separate whole cell proteome into biphasic layers. Total number of aqueous phase proteins, as assessed by 2DE, was comparable to the proteins isolated with commonly used detergent containing .. View More»
DOI:
10.4172/jpb.1000142