Editing ECL (section)

=== '''Two Solutions Method 2'''<br>  ===

Take two containers (they need to be something with a secure lid and which you can keep out of the light) and label one ‘A’ and one ‘B’. 

In the '''first tube (A)''', combine the following: 

1 ml luminol solution<br> 0.44 ml coumaric acid solution<br> 10 ml Tris-HCl 1M, pH 8.5<br> Distilled water up to a final volume of 100 ml 

In the '''second tube (B)''', combine the following: 

64 µl hydrogen peroxide<br> 10 ml Tris-HCl 1M, pH 8.5<br> Distilled water up to a final volume of 100 ml 

'''STORAGE''': Keep both solutions in the dark at 4°C. I used to wrap my solutions in aluminium foil to keep the light away from them, or keep them in a dark-coloured glass flask. 

'''SHELF LIFE''': The solution generally lasts up to 1-2 months if stored correctly. If you’re concerned that it’s been sitting in the fridge for too long, it’s probably best to just make more. 

NOTE: You can easily halve or quarter the volumes of solutions described here if you want less than 100 ml each of solutions A and B – if your lab only does western blots sporadically, this may be useful to you, as it’ll waste less solution if it goes off in between experiments.<br>