Multiple immunoreplica Technique: screening for specific proteins with a series of different antibodies using one polyacrylamide gel |
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Authors: | R P Legocki D P Verma |
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Institution: | Department of Biology, McGill University, Montreal, Canada H3A 1B1 |
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Abstract: | A simple method for immunological identification of proteins resolved electrophoretically is presented. Proteins from one polyacrylamide gel can be subjected to a series of electrophoretic transfers to nitrocellulose paper (partial “western-blots”), providing several replicas of the gel. Each replica can be reacted with a series of different antisera (at least three), where the preceding antibody is removed by treatment with pH 2.2. The antigen-antibody complexes are visualized using 125I-Protein A. Reactivity and antigenic specificity of proteins immobilized on nitrocellulose paper is not affected by repeated incubations and low pH treatments. Identical size of the replicas and superimposable profiles of proteins detected by antibodies allow a precise localization of particular polypeptides in the original gel. |
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Keywords: | To whom requests for reprints should be addressed |
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