Abstract:Three synthetic decapeptides of subunit Ⅳ of cytochrome b6-f complex were coulpled respectively with ovalumine and purified using High-Performance Liquid Chromatography. They were served as antigens and injected into rabbits. Five weeks later, three kinds of monoclonal antibodies were obtained. Western blotting showed that there were immunoaffinity reactions between anti 17 kD-1, anti 17 kD-2, anti 17 kD-3 and 17kD band of cytochrome b6-f complex from spinach chloroplasts. Enzyme-linked immunosorbent assay showed that there were immunoaffinity reactions between anti 17 kD-1, anti 17 kD-2, anti 17 kD-3 and thylakoid membranes from spinach chloroplasts. Therefore, these antibodies can be used as probes to survey the location and orientation of 17 kD on the two sides of thylakoid membranes.