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Enzyme-linked immuno-filtration assay applied to accelerated immunodetection of gel transferred proteins on
J M Pinon1, D Puygauthier-Toubas, H Lepan
1Laboratoire de Parasitologie-Mycologie, Hôpital Maison Blanche, Reims, France.
Abstract:
The enzyme-linked immunofiltration assay (ELIFA) has been used for the rapid detection of electrophoresed proteins transferred from gels to immobilizing matrices. By controlled filtration, ELIFA permits the saturation of nylon or nitrocellulose membranes and the immunodetection of blotted antigens to be carried out in 15 min. The method is simple, can be automated and requires no handling of membranes. It complements the well standardized steps of gel electrophoresis and semi-dry horizontal electroblotting which can themselves be carried out in less than an hour. The sensitivity is at least 1-5 ng. The same process can be extended to the accelerated characterization of glycoproteins using appropriate ligands, or to the identification of antigens in a variety of biological fluids.