Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications
A method has been designed for electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets. The method produces quantitative transfer of ribosomal protein gel containing urea. For sodium dodecyl sulfate gels, the original band pattern is obtained without loss of resolution, but the transfer is not quantitative. This method allows the detection of proteins by autoradiography and simpler than conventional procedures. The immobilized proteins were detected by immunological procedures. All additional binding capacity on the nitrocellulose was blocked with excess protein; then the specific antibody bound and, finally, a second antibody directed against the first antibody. The second antibody either radioactively labeled or fluorescein or peroxidase conjugate. Certain proteins are then detected by either autoradiography, under UV light, or with a peroxidase reaction product, respectively. In the latter case, as little as 100 pg of protein clearly d...