I have to do immunofluoroscence of pERK and p-p90RSK from CST on freezing microtome cut sections;paraformaldehyde fixed ;from mice brain(28-30 g mice)
- I plan to go for 4% PFA in PBS
- Post fixation 2hrs in same fixative
- Cryoprotect in 30% sucrose from 6-12 hrs(let me know the time for adult mice)
- 30% sucrose made in PBS; should i add ehthylene glycol for cutting on freezing microtome the brain slices
- How i store brains sections for a month
- should i go for floating method or on glass slides; which thickness would the best FOR THESE 2 PHOSPHO PROTEINS. any special suggestion for staining phosphoproteins
- Could plz correct if i am wrong in any way or plz give yr kind experienced suggestions
thanks
Hi Namratta,
here are answeres to your queries
Good luck and cheers for SSI