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The general workflow of the quantitative phosphoproteomics strategy for human spermatozoa samples analysis from sperm donors. (A) Purified sperm cells from semen samples (spermatozoa isolated from other cells and the seminal plasma). (B) Sperm protein extraction and digestion after cellular lysis. (C) Phosphopeptides enrichment and peptide separation. (D) The analysis and quantification of peptides. (E) The collection and analysis of sperm phosphoproteome data by bioinformatics tools.
