骨组织β-gal免疫染色实验技术方法
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- Dissect bone out of mice.
- Fix at tissue in fixative for 30min-2h at RT or overnight in 4�C with 4% paraformadehyde.
- Rinse with PBS.
- Soak tissue in X-gal staining solution for 4 hours at 37�C.
- Pour off the staining solution, replace with wash buffer, 2 times.
- Sock tissue in 30% sucrose in PBS.
- Embed and cut frozen section.
- Bone after fixed and briefly rinse with PBS.
- Soak with decal solution. Gently rock at 4�C for 1-3 days.
- Soak with 30% sucrose in PBS + 2mM MgCl2 at 4�C for overnight.
- Embed in OCT.
- Cut frozen section. Place slides in wash solution.
- Incubate sections with x-gal solution at 37�C for 1-3 hours (monitor staining every 30 min. It usually takes 2 hours).
- Rinse slides in PBS, 3 times.
- Rinse with 95% EtoH and stain with Eosin 20 secs for counterstain.
- Dehydrate and coverslip
