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From 1 to 28 days after frog sciatic nerve damage, dorsal root ganglia were incubated with [35S] methionine, and the labeled, rapidly transported proteins at various points along the nerve were analyzed on two-dimensional gels. The results show a dramatic increase in the labeling of a protein, which we have designated as A25, only after the arrival of the rapidly transported proteins at regenerating nerve tips. This effect is first seen 3 to 5 days after injury. On gels from regenerating nerves, A25 appears as a series of intense spots with an apparent molecular weight of 70,000. A25 is retrogradely transported from the regenerating nerve tip regions. Since labeled A25 increases only after the rapidly transported proteins reach regions of nerve containing regenerating axons, we conclude that it most likely arises from post-translational modification of a transported protein. Various experiments were conducted to rule out alternative sources of A25 labeling at the nerve periphery.