Immunostaining

Equilibrate slides to RT
Rehydrate in blocking buffer for 45min-1h in Coplin jar
Incubate with primary antibody in blocking buffer 4C O/N in humidified chamber
Use 170ul Ab/slide cover with hybrislips. Use water soaked Kimmies to create humidified chamber

Remove hybridslips
Wash 20min x3 with blocking buffer
Incubate with secondary antibody in blocking buffer 1h RT dark
Wash 20min x3 with blocking buffer
Mount slides using VectorShield + DAPI (1 drop/ slide) with glass coverslip
Seal with nail polish
Dry
Store at 4C dark

Blocking buffer: 1% heat-inactivated goat serum in PBS + 0.1% TritonX-100