gelatin staining with coomassie: how do I make it work?
Posted 11 April 2011 - 02:46 AM
I have been trying to set up gelatin zymography in my lab and ran into a strange problem. When I previously stained gels with a really old and dark Commassie solution somebody once gave me, the gelatin inthe SDS polyacrylamide gel stained nicely, giving a dark blue background over which I could see MMP digestion bands. When I ran out of this solution, I bought Bio-Rad Biosafe Coomassie, just to find out that no matter how long I incubate the gel with it, it will stain the protein bands but it never stains the background, making it impossible to discern any digestion bands...
Any suggestion as to why this happens? Why would gelatin not be stained by this type of Coomassie?! (I use 300 bloom gelatin at a final concentration of 1 mg/ml)
I am willing to buy a different Coomassie, but which one would be best to overcome this problem?
Posted 11 April 2011 - 09:58 PM
Posted 12 April 2011 - 01:07 AM
Posted 12 April 2011 - 08:32 AM
genius does what it must
i do what i get paid to do