Fig. 4. Final purification of the sperm chemoattractant by size-exclusion
chromatography. (A) The bioactive fraction from RP-HPLC was chromatographed on
a size-calibrated Bio-Sep S-2000 column, eluted at 1 ml min-1 in
water and absorbance monitored at 220 nm. Fractions corresponding to detected
peaks were pooled, lyophilized, resuspended in sea water, and bioassayed with
freshly spawned sperm. Numbers indicate the elution times of size standards
used to calibrate the column. (B) Sperm swimming response to isolated
fractions. Values are means + S.E.M. Fresh egg-conditioned sea water (ESW) and
filtered sea water (FSW) were used as positive and negative controls,
respectively. *Means significantly greater than negative controls
(P<0.0001). (C) Chemotactic response of abalone sperm to isolated
size fractions. Values are means + S.E.M.
