This study aimed to find a link between seminal plasma protein profile and semen characteristics, freezing ability and subsequent field fertility of bovine and buffalo bulls. Pre- and post-thawed semen of five Holstein and four buffalo bulls was used in this study. Total pattern of proteins precipitated from seminal plasma of bovine and buffalo bulls showed four different proteins of apparent molecular masses, 14 kDa, 15 kDa and 30 kDa with informative polymorphism in number and intensity of bands in both buffalo and cattle seminal plasma proteins. Seminal proteins bands (BSA-A1, A2, A3 and BSA-30) were positively correlated with percentages of motility and livability of spermatozoa, while negatively correlated with percentage of sperm abnormalities and acrosomal integrity in pre- and post-thawed semen of buffalo and bovine bulls. . It could be concluded that seminal plasma proteins can be used as a predicting measure for quality of bovine and buffalo semen. Further studies might be needed for quantitative determination of seminal plasma proteins which might give a better indicator for bull fertility when compared to the descriptive methods.