I am trying to Quantify Collagen Type I(1mg/ml) in a 5% Glucose pH 2.7 Solution with a 1:1 Ratio. I am trying to be very specific so any qualitative ways would not help. So Im ofcourse wanting to run a Gel. I have tried using a 1x, and 2X buffer solution in a Nupage 10% Gel, and got no Protein. In Fact after staining I can still see the Protein stain up top in the Gel. So i assumed it was a pH problem and tried a Tris pH 8.0 solution to first neutralize the solution and then run. Ive also tried using a 5X buffer and boiling the solution for about 30 minutes. It seams I cant get the Collagen type I to fully denature to run properly on a Gel. Any help would be great. I read up on some stuff here and seams like the Collagen is a pain to denature. but any help would be appreciated.
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