Chemiluminescent Detection¶
Contributed by Ian Chin-Sang, Queens University, ON, Canada
Homemade chemiluminescent detection recipe
Requirements¶
p-coumaric acid (Sigma #9008) 3-aminophtalhydrazide (Luminol, Sigma #09253) 1 M Tris pH 8.5 30 % H2O2 90mM p-coumaric acid in DMSO (store at -20°C) 250 mM Luminol in DMSO (store at -20°C)
Method¶
- Solution A (50ml):
- 5 ml 1M Tris pH 8.5
- 45 ml water
- 110 ul 90 mM coumaric acid (0.2mM coumaric acid)
- 250 ul 250 mM Luminol (final 1.25mM Luminol)
Keep 2-3 weeks at 4°C
- Solution B (1ml):
- 100 ul 30% H2O2
- 900 ul water
Keep 2-3 weeks at 4°C
- Combine 1 ml Solution A + 3 ul Solution B (5ml Solution A +15ul Solution B for each blot)
This method is based, with permission, on an original protocol available here.