Proteomic analysis of any cell type is highly dependent on both purity and pre-fractionation of the starting material in order to de-complexify the sample prior to liquid chromatography mass spectrometry (MS). By using back-flushing techniques, pure spermatozoa can be obtained from rodents. Following digestion, phosphopeptides can be enriched using TiO2.
Baker, M. A., Hetherington, L., Weinberg, A., Velkov, T. Phosphopeptide Analysis of Rodent Epididymal Spermatozoa. J. Vis. Exp. (94), e51546, doi:10.3791/51546 (2014).