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Chapters
- 00:05Title
- 01:34Preparation of Cannula
- 02:43Removal and Cannulating of Mouse Epididymis
- 03:47Backflushing, Washing, and Lysing of Caudal Epididymal Spermatozoa
- 05:09Disulfide Bond Reduction, Alkylation, and Precipitation
- 06:12Trypsin Digestion and Phosphopeptide Enrichment
- 07:32Results: Spermatazoa Isolation from Rodent Epididymis
- 08:50Conclusion
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.
