Sperm protamine-1 (P1) and protamine-2 (P2) concentrations were evaluated concomitantly with global DNA methylation patterns in a population of male infertility patients. Protamine quantification and immunofluorescence microscopy in conjunction with quantitative image analysis revealed no significant relationships between the P1/P2 ratio, P1 concentrations, or P2 concentrations and levels of global DNA methylation in mature spermatozoa.