A Novel Approach for the Analysis of Membrane Proteins Applied to Glioma Stem Cell Xenografts

摘要

In this study we improved the identification of membrane proteins through 1) optimized subcellular fractionation, 2) increasing organic solvent in the digest, and 3) LC-MS loading buffer with urea. Keeping hydrophobic membrane peptides soluble in the autosampler (4°C) with chaotropes improves sample pickup, column loading, and protein identification with minimal carbamylation of peptides, which negatively impacts label-free quantification. Test other reagents that may improve peptide solubility for autosampler pickup and subsequent column loading and LC-MS analysis. Improve number of identifications with 2D-LC-MS/MS.