Stock solutions (-20°C)

  • X-Gluc (5-bromo-4-chloro-3-indolyl-β-D-glucuronide Cyclohexylammonium salt)  20 mg/m dissolved in DMF
  • 500 mM Phosphate buffer (pH 7.2)
  • 12.5 mM K3Fe(CN)6　
  • 12.5 mM K4Fe(CN)6
  • Triton X-100 (Conc)
  1. GUS buffer (For 10 ml)
  • 1 ml  500 mM Phosphate buffer (pH 7.2)   final 50 mM
  • 0.4 ml 12.5 mM K3Fe(CN)6  final 0.5 mM
  • 0.4 ml12.5 mM K4Fe(CN)6  final 0.5 mM
  1. GUS staining buffer (For 4.1 ml)
  • 0.1 ml 20 mg/mL X-Gluc
  • 4 ml GUS buffer
  • 4 µl Triton X-100
  1. Immerse plant samples in 1 ml of 90% cold acetone for 10~15 min in microtube
  2. Discard acetone and wash samples with GUS buffer (1 ml) twice
  3. Add 0,5 ml GUS staining buffer and vacuum infiltrate until samples were sedimented
  4. Incubate samples at 37°C for o/n
  5. Rinse samples with 70% EtOH twice