Parvalbumins are highly stable Ca2+ binding proteins, present in large quantities in the sarcoplasmic reticulum in the white muscle of most lower vertebrates and fish. The properties of these proteins make them promising antigens for the use as a specific biomarker for fish species identification. Parvalbumin isotypes were isolated, on a preparative scale level, by use of size exclusion chromatography (SEC) and anion exchange HPLC. The utility of this technique, with regard to maximizing purified isotypes, is discussed.