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Experimental Protocols
Embryo Medium
Embryonic Ganglia
Luria Broth
50x TAE
Antibody Staining
Pond Water
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Antibody Staining of Fixed Whole Embryos
- Ansthesize in 6-8% EtOH in Wenning's 2 min RT.
- Fix 1 hour RT. Rinse in PBS 2x.
- Quench
- Permeabilize PBS/1% TX 100 4X 30 mins each. Gentle shaking. 2hrs
- Block 2% NGS of 2nd Ab species. In PBS 0.2% TX 100 (or 5% milk) 1 hour RT or O/N 4.C.
- 1st Ab. In block 1/10. 1/5000 4hr or O/N 4.C.
- Wash PBS 0.2% TX 100 5X RT shaking.
- Block again
- 2nd Ab in Block 3 hr or O/N 4.C.
- Wash PBS TX
- Place in desending 90, 85, 70% EtOH (EtOH only with HRP or AP staining, with fluorscent staining go directly to 50% glycerol in water then
coverslip, nail polish around edge to keep from drying out).
- Mount in gelvatol, coverslip Paramount.
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