Victor, Entry #3: Western Blots

Hi everyone,

Quantifying the protein level before the western

I hope everyone is having an interesting and enjoyable summer. I just finished my fifth week here in Baltimore in my lab that focuses on early detection of pancreatic cancer. Now that I have been here for five weeks they are starting to trust me a little more. In addition to helping out with the CAPS study as I have mentioned in my previous posts they are asking me to run experiments like western blots and DNA isolation while giving me more independence. This transformation has been one that I am very happy about. One of the projects that I am helping out with involves biomarker research. Finding a clinically applicable biomarker of pancreatic cancer has been the aim of early detection cancer researchers for a while now. Finding one would have enormous implications.as current diagnostic tests are invasive and rather expensive. A biomarker would drastically reduce the price of diagnostic tests so then diagnostic tests could be readily administered to not only people who are at risk for pancreatic cancer but also the general population. This would cause the prognosis of pancreatic cancer to improve drastically because more pancreatic cancers would be caught earlier in the development when the lesions are more likely to be resectable and less likely to have metastasized to other parts of the body. However, the search for a reliable biomarker has proven to be extraordinarily difficult. One of the side projects that a postdoc in my lab is working on is researching the biomarker potential of the protein mesothelin. Mesothelin is a membrane protein that is known to be over expressed in tumor cells but more research needs to be done in order to determine if it could potentially be used as a biomarker. The hope is that this protein could be used to detect cancerous exosomes within the blood In order to examine the protein expression in tumor cell lines a western blot was preformed.

Results of the blot show some expression although it is somewhat faint 

The results of the blot (depicted above) were somewhat faint and had to the film had to be exposed for over 40 minutes. We suspect it has to do with the antibodies but we are not sure because there are so many things that could have gone slightly wrong.