Comparison of protein precipitation methods for sample preparation prior to proteomic analysis.
Protein samples should be free of salt and other disturbing agents and have an appropriate concentration to be suitable for two-dimensional (2D) electrophoresis, the principal step of proteomics. To find the most efficient method for sample preparation, we used human plasma and compared four widely applied precipitation methods, using trichloroacetic acid (TCA), acetone, chloroform/methanol and ammonium sulfate, as well as ultrafiltration. Precipitation with TCA and acetone and ultrafiltration resulted in an efficient sample concentration and desalting. We also found that ammonium sulfate fractionation can efficiently remove albumin, which represents more than 50% of plasma proteins.
Jiang
Lei
L
Bio-X Life Science Research Center, Shanghai Jiao Tong University, Hao Ran Building, P.O. Box 501, 1954 Hua Shan Road, Shanghai 200030, PR China.
He
Lin
L
Fountoulakis
Michael
M
D000645:Ammonium Sulfate / Q000737:chemistry; D001798:Blood Proteins / Q000737:chemistry / Q000302:isolation & purification*; D011232:Chemical Precipitation; D015180:Electrophoresis, Gel, Two-Dimensional; D020543:Proteome
D001798:Blood Proteins; D020543:Proteome; D000645:Ammonium Sulfate