Guinea pig anti doublecortin dcx

For the detection of migrating immature neurons or neuroblasts within the SVZ sections were fixed in 4% paraformaldehyde (PFA in 0.1 M PBS, pH 7.4, for 15 min RT) and then washed in PBS (0.1 M; 3×5 min) prior to antigen retrieval with 10% proteinase K (DAKO, Glostrup, Denmark) in 0.1 M PBS for 3 min following by incubation in 30% fetal calf serum to stop proteinase K activity for 30 min. Sections were washed in PBS (0.1 M; 3×5 min) followed by incubation in blocking solution containing 5% NGS in 0.1 M PBS for 30 min. Sections were then incubated with guinea-pig anti-doublecortin (DCX; 1∶3000, Millipore, Billerica, MA, USA) for 1 hour in a mixture of PBS (0.1 M; pH 7.4) containing 5% NGS prior to PBS washes (0.1 M; 3×5 min) and incubated with secondary antibody biotinylated goat anti-guinea-pig (1∶200, Vector Labs, Burlingame, CA, USA) for 30 min. Sections were further analysed as described above.