Collagen II western blotting - (Jul/06/2010 )

Hi Everyone...

Could you please advise me on how to check for the presence of collagen II in my cell layer samples? I dont know if I should check the medium or the cells for the protein. I have purchased this antibody MAB8887 from chemicon (millipore) and it says that In Western blotting, MAB8887 reacts with the TCA fragment of lathyritic type II collagen after digestion with mammalian collagenase. It also reacts with pepsin-digested type II collagen. It will not react with intact Collagen II under traditional conditions.
I have no idea what collagenase type I should use.....
Further, I do not know how to do pepsin digest either.
If anyone would know how this can be solved, please please help me... there is just no body in my lab workin on this...and I am helpless...

scarlet

Would there happen to be any related research papers to your work? Most papers have a pretty detailed methods section where they describe their processes.

Apart from that, I work with tissue samples.. so am not much help =/