Paraffin embedded formalin-fixed tissue sections from rat brain and patient material were placed in xylene bath for 263 minutes, absolute ethanol 263 minutes, 96% ethanol 262 minutes and finally in distilled water for 30 seconds for removal of paraffin and rehydration. Epitope retrieval was performed by heating the sections at 99uC for 20 minutes in 10 mM citrate buffer at pH 6.0. The sections were incubated with a monoclonal human-specific anti-nestin antibody 1:200 in TBS/1%BSA over night at 4uC. A biotinylated goat-anti-mouse antibody (Vector Laboratories) was used as secondary antibody (dilution 1:100) for 1 h at room temperature followed by ABC-complex incubation for 30 min. Sections were developed with with 393-diaminobenzidine (DAKO Cytomation), following the manufacturer's instructions.