Mixed Protein Solution For Pancreatic Cancer

1330 Words 6 Pages
Currently, pancreatic cancer is the fourth leading cause of cancer-related deaths in the United States. Furthermore, the five year survival rate of individuals diagnosed with pancreatic cancer has been estimated at just below 5%. This can be attributed to the fact that pancreatic cancer often progresses stealthily and physical symptoms (jaundice, abdominal pain, and weight loss) do not present until the cancer is in its later stages. As a result, the best way of treating pancreatic cancer is to detect and treat it in its early stages. With that being said, there have been no real advances in the early detection of pancreatic cancer. Most current methods of detection, such as biopsies, are too invasive and costly. However, recent research …show more content…
The proteins from each solution will then be precipitated by ethanol and suspended in 2-D cell lysis buffer (). Next, the proteins from the healthy volunteers and the sick volunteers will be stained with CyDye Cy3 and Cy5, respectively, and combined into one solution. The resulting solution will then be subjected to 2-dimensional gel electrophoresis. This will be done by transferring the mixed protein solution to isoelectric tube gel (pH 3-10) and subjecting it to a constant electric current (145mA). After the gel has been allowed to run for about an hour, the isoelectric tube will be rinsed with running buffer and set aside. Next, an SDS gel will be prepared in order to separate the proteins based on mass. This will be done by sandwiching two spacers between two glass plates and placing them within a plastic bag. The plate/bag assembly will then be placed into a casting stand and filled with a solution consisting of 12% gel mix, 80 microliters of 100 mg/ml ammonium persulfate, and 15 microliters of TEMED. Next, a comb will be wedged between the two plates and the solution will be allowed to solidify. Once solid, the gel will be removed from the plate/bag assembly and placed vertically in an electrophoresis apparatus. Afterwards, the isoelectric tube gel will be placed horizontally on the sds-gel and 1X running buffer will be poured into the upper and lower apparatus chambers. The gel apparatus will then be connected to a constant voltage source (175v) for 1 hour, allowing the proteins within the tube to segregate based on mass. Once separated, software will be used to scan the 2-dimensional gel and determine the fold change of the protein expression

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