I am doing a Western blot of an isoform of fibronectin (EDB) that is present in cancer. Normally, I would use beta-actin as a loading control. However, fibronectin is secreted into the matrix, so I have collected protein from the media. Therefore, actin will not be there. I cannot use another protein that is highly expressed in cancer, such as collagen, as a control and I am still learning about the composition of the extra-cellular matrix. So, I was planning to do some research, but I thought I would ask here too, in case anyone has done any similar work. Thanks for your help!
Cytochrome-C. It shows equal levels as far as there is no cell death in the system. Refer Supplemental figure-3 of my paper
http://www.nature.com/articles/cddiscovery20163
Hi Sarah, when you are doing western blots of cell extracts or secreted proteins the best control is to load equal amounts of protein.
Dear Sarah
Fibronectin (FN) is a ubiquitously expressed, multi-domain glycoprotein that is critical for development and that is instrumental in numerous physiologic and pathologic processes.
For more information please check the following publication: CONTROL OF FIBROBLAST FIBRONECTIN EXPRESSION AND ALTERNATIVE SPLICING VIA THE PI3K/AKT/mTOR PATHWAY. Exp Cell Res. (2010)1; 316(16): 2644–2653.
Hoping to be helpful
Marius
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