Evaluation of Sample Preparation Methods for Label-Free Quantitative Proteomics of Human Brain Tissue

摘要

Sample clean-up with C18 spin columns increased protein IDs by 10%. Addition of RapiGest for solubilization did not increase protein IDs. Independently analyzed biological replicates have 72%-75% overlap in protein IDs. Lysis in 8M urea buffered with 1M ammonium bicarbonate (in-solution) and 5% SDS (in-gel) are comparable and both function for label-free quantitative proteomics studies of post mortem human brain tissue. 5% SDS is preferable due to the low viscosity of the resulting lysate.