Coating and seeding: 22.05.18 [see the post 20180522 – 20180529: HTB-11 cells diferentiation (exp. 4) + ICC]
ICC protocol:
- Fix with PFA 4% (aprox 5 ml/plate). Incubation at RT during 10min.
- Wash with PBS 1x
- Blocking solution: Goat Serum 5 % in PBS for 1h at RT.
- Primary antibody: anti-neurofilament -L clone EP675Y, rabbit monoclonal antibody (04-1112, Millipore). Diluted 1/500
- Wash with PBS 1x
- Secondary antibody: GAR488 IgG diluted 1/1000
- Wash with PBS 1x
- Vectashield mounting medium
Results:
- There are few axons. Most of them are detached from the coverslip. Change the fixation method.
