Mouse monoclonal anti pcna

After BLI imaging, brains were washed with ice cold PBS and post-fixed in 10% formalin for 72 hours and embedded in paraffin blocks. 5μm coronal sections were used for all histological studies. All IHC procedures were performed as previously described (Becher et al. 2008 (link)). The following antibodies were used at the stated dilutions: rabbit polyclonal anti-Iba1, 1:100 (Wako Pure Chemicals, Osaka Japan); rabbit polyclonal anti-GFAP, 1:8000 (DAKO, Denmark); rabbit polyclonal anti-Olig2, 1:250 (Chemicon, Temecula CA); mouse monoclonal anti-PCNA, 1:2000 (DAKO, Denmark), mouse monoclonal anti-NeuN, 1:50 (Millipore) and goat polyclonal anti-SHH, 1:100 (Santa Cruz N-19). For IHC detection, an automated staining processor was used (Discovery, Ventana Medical Systems, Inc.) with protocols developed in the Holland and Hambardzumyan laboratories. For immunofluorescence (IF) staining, secondary antibodies conjugated to different Alexa-Fluor dyes (488, 555, 647 from Invitrogen) at a dilution of 1:500 in PBS/2%BSA were applied. For nuclear counterstaining, DAPI was used (Sigma).