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Fig. 8. Proteins isolated from the basic region of an IEF gel containing ovulated zona pellucida (ZP) glycoproteins inhibit wild-type and GalT I-null sperm binding to ovulated eggs. Proteins were isolated from acidic (fraction 1), neutral (fractions 2 and 3) and basic (fraction 4) regions of an IEF gel containing 5000 ovulated ZP and tested for biological activity in the sperm-egg binding assay. The acidic fraction (containing ZP3) inhibits wild-type, but not GalT I-null, sperm from binding to ovulated eggs. By contrast, the basic fraction (containing the basic ligand) prevents both wild-type and GalT I-null sperm from binding to ovulated eggs. Doubling the amount of starting material lead to 69% inhibition of sperm-egg binding by fraction 4. Each bar represents the mean±s.e.m.; n=4 experiments. In a single experiment, the value for each bar represents the average of three determinations (for a total of 12 assays per bar). The broken line indicates the number of sperm bound in the control, i.e. material obtained from a blank IEF gel treated identically to gels containing zona glycoproteins.





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