Purification of pig serum haemopexin by haemin-Sepharose affinity chromatography

Haemopexin was isolated from pig serum in pure form by affinity chromatography and ion-exchange chromatography. The affinity gel synthesized contained about 0.3 mumol/ml haemin covalently linked to AH-Sepharose 4B. The molecular weight of the protein was measured by polyacrylamide gel electrophoresis in the presence of SDS followed by sensitive silver staining. This procedure and also immunoelectrophoretic studies indicated purity. The mobility of haemopexin in polyacrylamide gels changed following reduction with dithiothreitol.