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System

Illustration

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

  1. Place dissected adult tissues in PBS and cut them with a blade.
  2. Transfer tissues in small dishes or multi-well plates.
  3. Fix with the fixative solution for 30 minutes.
  4. Rinse tissues with 1%Triton X-100/PBS 10minutes.
  5. Rinse tissues with 2 changes of PBS for 10minutes each.
  6. Incubate tissues in X-gal working solution overnight.
  7. Post-fix with the fixative solution for 30 minutes.
  8. Rinse with 2 changes of PBS for 10minutes each.

Note: Tissues were rocked gently at room temperature on each step.