Today we did not have any planned activities, so the students planned outings for the day. A large group of students, myself included, went to Palermo via subway with Hannah. Once we got to the neighborhood, Hannah allowed us to walk around on our own and told us to be back at the hostel by 3 o'clock to pack.
To prepare for tomorrow's Intel Northwest Science Expo, I met with Aline yesterday to go over my presentation and hear her feedback. I printed the poster to the size of an 8x11 piece of paper. First, she read over the poster, and then asked me questions about it. She said my poster is very clear and that there is a nice balance between graphs/diagrams and text. Then, I went through my whole presentation, inviting her to ask questions as I went through the poster.
After running the gel, transferring the gel, blocking, and probing with primary antibody, I washed the primary antibodies with TBST, then probed with secondary antibody corresponding to whether it was anti-mouse or anti-rabbit (see below). Afterwards, the western blots were imaged.
Preparing for another western blot: From the Bradford assay of the BaF3 serum-free (SF) lysates, I calculated 50ug/ul per well (for each sample) and enough for 5 gels. With the protein concentration calculated, I determined how much lysis buffer to add up to a volume as well how much sample buffer (dye) to put in each sample. I kept these samples in the -80 freezer.
Last Thursday, I lysed the 293T17 cells. Since the media had already collected virus, I also kept the media.
Today, after a quick breakfast and complicated commute, we arrived at ESMA, a former naval academy and perhaps the most infamous detention center of the southern cone. Despite our punctual arrival just before our scheduled tour at 10:00, we were informed that unforeseen consequences had led to the postponement of our tour until 2:00.