Hi,
I want to evaluate the expression of 6 proteins (3 receptor and 3 enzyme) in rat vessels which has a diameter of 150-300 um. I will do western blotting and qPCR for this purpose. Hovewer both vascular smooth muscle cell and endothelial cells have expression of 6 proteins. So i want to know the change in amount of proteins in endothelium. How can i solve this problem? Which method do you suggest? I thought I can show and quantify the change with immunohistochemistry but i am not sure. Please help me.
Thanks.
You can extract the blood vessel from mouse. Digest the vessel with collagenase to make single cell suspension. After that, you need to sort CD31 positive cell and isolate RNA from these cells.
Thanks a lot. It is very clear answer. What do you think about immunohistochemistry?
You can do immunohistobiochemistry as well. In that case you need to look for co-localisation of CD31 and your protein of interest.
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