DETECTION OF DNA-BINDING PROTEINS AFTER ELECTROPHORESIS ON DENATURING POLYACRYLAMIDE GELS

A simple method is presented for detecting DNA-binding proteins after separating them on polyacrylamide gels containing sodium dodecylsulfate. The separating gel contains radioactively labelled DNA and, after electrophoresis, is washed in buffer to remove sodium dodecylsulfate and allow renaturation of proteins. DNA.is washed out except where it is retained by proteins which regain their DNA-binding properties. The proteins can be visualized by autoradiography and appear as black bands on a shaded background after one-dimensional electrophoresis, or as black spots after two-dimensional electrophoresis. Compared with the standard method, the new method exhibits high sensitivity with rat liver protein fractions.