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A bioassay for the sperm attractant from Oedogonium cardiacum was developed. The attractant was confirmed to be highly water soluble, destroyed by heating to 100 C and by dilute acid or alkali. It was fractionated by gel filtration with the attractant accompanied by yellow pigments. It has a molecular weight between 500 and about 1500, based on gel filtration, ultrafiltration, and dialysis. Freezing changes this to both larger and smaller molecules. It appears to have cationic groups. A paper chromatographic separation was found that divided the pigment into at least 3 major bands, one of which was closely associated with the attractant. An absorption spectrum of this pigment showed 1 peak in the visible spectrum at 417 mμ and 3 well-defined peaks in the ultraviolet region. The lack of a peak at 280 mμ indicates the attractant is not a polypeptide. [ABSTRACT FROM AUTHOR]