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 detectable. It is anticipated that the procedure will be applicable to the analysis of various proteins with a particular reaction or a ligand.

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1971 initial criteria for the classification of systemic lupus erythematosus (SLE) is revised and updated to incorporate new knowledge and enhance immunological disease classification. 1982 revised criteria including fluorescence antinuclear antibodies and antibodies to native DNA and Sm antigens. Some criteria involving the same organ system are collected into a single criterion. Raynaud's phenomenon and alopecia are not included in the 1982 revised criteria for sensitivity and low specificity. New criteria was 96% sensitive and 96% specific when tested with SLE and control data collected from 18 patients participating clinics. When compared with the 1971 criteria, 1982 revised criteria showed gains in sensitivity and specificity.

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