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The dye 1-anilino-naphthalene-8-sulphonate (ANS) bound at 94.5 +/- 2.7 pmol per 10(6) spermatozoa (KD = 1.83 +/- 0.13 X 10(-5) M) and 7.30 +/- 0.17 pmol per mg seminal plasma protein (KD = 2.19 +/- 0.06 X 10(-5) M). Equilibration of excess ANS with a mixture of washed spermatozoa and seminal plasma resulted in a significant increase in fluorescence over that calculated by summation of the individual levels (P less than 0.05). A less pronounced but significant increase was seen when an egg-yolk lipoprotein, previously shown to have cryoprotective properties, was similarly added to washed spermatozoa (P less than 0.05). This increase in fluorescence was not reduced by washing spermatozoa--lipoprotein mixtures, suggesting that an interaction between the lipoprotein and the cell membrane had occurred.

reproduction

A spectrofluorometric investigation, using 1-anilino-naphthalene-8-sulphonate, of the interaction between washed bovine spermatozoa and seminal plasma or egg-yolk lipoprotein