Last Western Blots

This week I began my second round of Western Blot tests. The first test I completed was to determine the proper conditions to do the second test under. I found that it was best to apply no heat to the samples. I then went through typical Western Blot procedures. This time the process went more smoothly and I did not encounter any significant issues. On Tuesday I finally had my results. I took the elevator down to the second floor and placed my samples in the imager. After I got the results, I printed them out and went back up to the lab to get the help of the graduate students with interpretation.

I discovered two things from my test results. First, that the fourth tissue sample had not been properly refrigerated at some point in the past and was no longer viable. Second, that NASH has no effect on the transporter protein Mct8. NASH is known to disable many proteins within the liver and cause issues, specifically with drug metabolism. My Western Blot showed that Mct8 is not one of the proteins causing drug overdoses in people affected by NASH. This means that Mct8 can be ruled out and no longer considered. There were 13 tissue samples, three from a control rat with the protein Bcrp, three from a rat with Bcrp and NASH, three from a control rat without Bcrp, and four from a rat without Bcrp with NASH. All of these samples had Mct8 levels that were not significantly different.

After completing my first successful pair of Western Blot tests, I helped one of the graduate students in the lab with her Western Blot on Thursday. She had a meeting to attend so I handled the time sensitive parts of the test. It felt good to be getting proficient at this test and confident in my own abilities to complete every step correctly. I was not able to view the results of this test because the graduate students finished it on Friday but I was glad to help out. Next week I will begin focusing on my final presentation and my final product, which will be a conference style poster.