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[Aris]

Hi all,

I am doing experiments with endothelial cells and specifically I am looking at Collagen IV and Fibronectin expression levels as produced by the endothelial cells by Western Blot. Recently, I have been getting some strange thoughts that the lysis buffer that I am using is not potent enough to solubilise Collagen IV and Fibronectin. My lysis buffer is a Tris based buffer containing Triton X 100 and EDTA and I add a mix of proteinase inhibitors just prior to lysis. I am aware that in older times (10-15 ago) scientists used to melt basement membrane in order to solubise its components such as Collagen IV and Fibronectin.
Is there anyone out there that has more insight into this or a buffer to suggest or even a suitable publication that I can use as a guide?

Thanks a lot