Application of label-free proteomics for differential analysis of lung carcinoma cell line A549.
A label-free solution basing on a highly reproducible and stable LC-MS/MS system allows quantitative proteome analyses. Due to nonlabeling approach, the label-free method has the potential to measure samples from clinical specimen monitoring and comparing thousands of proteins. The presented label-free workflow includes in-solution digest, LC-MS analyses, data evaluation by the means of Progenesis™ software, and validation of the differential proteins. We successfully applied this workflow in a proteomics study analyzing the human lung carcinoma cell line A549 treated with transforming growth factor beta 1, a cell culture model of lung fibrosis. The differential analysis of only 1 μg protein per sample led to 202 significantly regulated proteins.
Department of Medical Proteomics/Bioanalytics, Medizinisches Proteom-Center, Ruhr-Universität Bochum, Bochum, Germany. barbara.sitek@rub.de
Barbara
B
Sitek
Daniel M
DM
Waldera-Lupa
Gereon
G
Poschmann
Helmut E
HE
Meyer
Kai
K
Stühler
D002457:Cell Extracts / Q000737:chemistry / Q000302:isolation & purification; D045744:Cell Line, Tumor; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D003627:Data Interpretation, Statistical; D015180:Electrophoresis, Gel, Two-Dimensional; D006801:Humans; D008175:Lung Neoplasms; D010449:Peptide Mapping; D059748:Proteolysis; D020543:Proteome / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D040901:Proteomics; D012984:Software; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D053773:Transforming Growth Factor beta1 / Q000502:physiology; D014357:Trypsin / Q000737:chemistry
D002457:Cell Extracts; D020543:Proteome; D053773:Transforming Growth Factor beta1; D014357:Trypsin