Shotgun Proteomics Analysis of Estrogen Effects in the Uterus Using Two-Dimensional Liquid Chromatography and Tandem Mass Spectrometry.
Shotgun (gel-free) proteomics is a useful approach to perform identification and relative quantification of protein in complex mixtures such as tissue homogenates, biological fluids, cell lysates, and extracellular proteins. Incorporation of separative and analytical techniques such as two-dimensional liquid chromatography at nanoscale (2D-nanoLC) coupled to tandem mass spectrometry (MS/MS analysis) into the shotgun protocol provides an excellent strategy. This chapter describes the application of the shotgun proteomics protocol to evaluate the identity and expression analysis of proteins from rat uterus after estrogen (ethinylestradiol) treatment. The steps of the protocol involve sample preparation (digestion), 2D-nanoLC-MS/MS analysis, and shotgun proteomics analysis including bioinformatics tools for data conversion, organization, and interpretation.
['Callegari|Eduardo A|EA|']
D000818:Animals D002853:Chromatography, Liquid D019295:Computational Biology D030562:Databases, Protein D004967:Estrogens D004997:Ethinyl Estradiol D005260:Female D036103:Nanotechnology D011506:Proteins D040901:Proteomics D051381:Rats D053719:Tandem Mass Spectrometry D014599:Uterus D057188:Workflow
Proteomics Estrogen Uterus Liquid chromatography Mass spectrometry Proteins Rat Peptides “Shotgun” proteomics