The Chromosome-Centric Human Proteome Project (C-HPP) aims at finding existence evidence of missing proteins (MPs) at the early stage. Based on previous research, it has been proved that human testis tissue is suitable for further research of MPs because of its high gene expression level and high transcriptome complexity. However, the limits of dynamic range in mass spectrometric analysis and the deficiency in sensitivity and resolution will seriously affect the detection efficiency. In this study, we used Q-Exactive HF to detect three individual testis tissues dealted with two separation methods (tricine-SDS-PAGE and SDS-PAGE).