We raised polyclonal antibodies against the C-terminal peptides of protein kinase C (PkC) subspecies alpha, beta 1, beta 2, gamma, delta, epsilon, and zeta and checked their specificity against brain extracts using Western immunoblot analysis. With equal amounts of protein applied to gels PkC subspecies beta 1, delta, epsilon and zeta were detected in primary cultures of mixed glial cells: bands for the alpha and beta 2 subspecies were less prominent. PkC gamma was not detected in primary glial cultures. The epsilon and zeta subspecies of PkC were detected in subcultures of type 1 astrocytes with weaker bands for the alpha, beta 1 and beta 2 subspecies. Blots of O-2A-lineage glia contained PkCs delta and zeta as prominent bands: the alpha, beta 1 and epsilon subspecies were also present. All PkC subspecies including PkC gamma were detected in C6 glioma cells.