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System
- The Exchangeable Gene Trap System
- Trap Vector
- Exchangeable System
- ES Cell Lines
- Isolation of Trap Clones
- Registration in EGTC
- Establishment of Trap Mouse Lines
- Deposition of Trap Lines in CARD
- Expression Analysis by X-gal Staining
- Reference
Expression Analysis by X-gal Staining
Description
X-gal staining was performed for detecting β-galactocidase reporter expression in a wide variety of adult tissues of each trap line.
X-gal Staining Procedure
Materials Fixative Solution
2.7% formalin
0.2% glutaraldehyde
0.02% Nonidet P-40
in PBS
X-gal Stock Solution
2% X-gal (5-Bromo-4-chloro-3-indolyl-beta-D-galactopyranoside) in
dimethylformamide
Store at -20℃ in small aliquots.
X-gal Working Solution
5mM Potassium ferricyanide
5mM Potassium ferrocyanide
0.5mM MgCl2
0.05% X-gal stock solution
in PBS
Make fresh just before use.
Procedure
- Place dissected adult tissues in PBS and cut them with a blade.
- Transfer tissues in small dishes or multi-well plates.
- Fix with the fixative solution for 30 minutes.
- Rinse tissues with 1%Triton X-100/PBS 10minutes.
- Rinse tissues with 2 changes of PBS for 10minutes each.
- Incubate tissues in X-gal working solution overnight.
- Post-fix with the fixative solution for 30 minutes.
- Rinse with 2 changes of PBS for 10minutes each.
Note: Tissues were rocked gently at room temperature on each step.