BRADFORD reagent - (Apr/04/2006 )
I did a Bradford reagent, but its color was dark blue instead of light brown (the right color)!!
I really don't know what I did wrong. The protocol that I followed is:
Dissolve 100 mg Coomassie Brilliant Blue G-250 in 50 ml 95% ethanol, add 100 ml 85% (w/v) phosphoric acid. Dilute to 1 liter when the dye has completely dissolved, and filter through Whatman #1 paper just before use.
I diluted to 1 liter with destiled water, is it right?
Can be pH the problem?
Please help me.
your glassware (plasticware?) may be contaminated.
What's wrong with buying pre-made Bradford's reagent?
100 mg of Comassie
50 ml of 95% ethanol
100 ml of phosphoric acid (85%)
+ 50 ml ddH20
That is 5 x Bradford reagent
I sometimes get a lightly greenish brown color that gets fully brown after adding a bit more phosphoric acid. If yours is dark blue it could be because of detergents in plastic/glassware.