Hello, is there anybody currently working on cell-secreted protease which is mainly focused on Matrix Metalloproteinase 9?
I'm new to cell manipulation research. I'd like to ask whether we should purify Metalloproteinase 9 (MMP-9) after being activated by p-aminophenylmercuric acetate?
I'm working to figure out the catalytic activity of MMP-9 on a customed peptide and I need to make sure the proenzyme form is totally converted into active form (My proenzyme is from Sigma). Is it necessary to add p-aminophenylmercuric acetate to activate the cell-secreted MMP-9, too?
Thank you for reading,
The APMA need not be dialyzed away after the activation of MMP. The solution can be used as such.
Cell secreted MMPs too can be activated using APMA.
Hi Diem, you have to activate the pro-MMP-9 with APMA before doing activity assays. And as Sangeetha said, you don't have to remove the APMA for your peptide assay.
If you are using immobilized gelatin to purify MMP-9, you can APMA activate it either before or after this step.
The best way to determine whether you have a preparation of activated MMP-9 is to run a gelatin zymogram with control preform.
To Dr. Sangeetha and Dr. Stefansson:Thank you so much for your valuable advices!
- Badges
- Science topic