Immunopanning of retrograde-labeled corticospinal motor neurons from early postnatal rodents

Here we describe a method to purify corticospinal motor neurons (CSMNs). It combines the versatility of retrograde labeling with the advantages of immunopanning. Rat cortices with CSMNs that have been labeled with cholera toxin beta (CTB) are dissected and dissociated, and the CSMNs are specifically purified via immunopanning with an anti-CTB antibody. We show the efficacy of this method in early rat pups by purifying CSMNs to >99% purity. The method can be used to highly purify any neuronal cell type whose projections can be selectively labeled via retrograde tracing.