Rotation #2 Diary: Week 3

Finished my third week of the rotation. My IP experiments... kind of worked. I was able to detect a faint band, not significantly better than just doing a regular western blot. However, doing the IP does make the blot come out a lot cleaner, so I'm going to try again next week. I'll tinker with the protein concentration and antibody concentration a little bit to try to maximize the amount of my desired protein in the end. And, I'll try using rat protein lysates instead of mouse protein lysates, since the antibody we're using was raised against the human protein, which has a greater degree of homology with rat than with mouse. 

Even if I can't get these blots to work, it's possible that the antibody may be able to detect my protein in histological specimens. If the blots fail me now, I will probably move on to immunohistochemistry to see what I can see. I've done a lot of that in the past. I have to say, though, I enjoy the precision of the more purely molecular techniques. Messing around with little pieces of tissue and mounting them on a slide is more annoying than working with nitrocellulose membranes. Although, I think for an ideal project I'd be doing a little of both -- studying protein/protein interactions, but also studying anatomical characteristics of the protein (or with mutations of that protein). Maybe even doing some behavioral studies or something, for the big picture. It's rewarding to get a super clean result in molecular biology, but it's also rewarding to actually see what's going on in an animal, either by looking at a representative brain section or observing a behavioral phenotype. Or doing physiology, I suppose, although I have absolutely no experience in that, and it seems like most e-phys people specialize in doing that and little else, because the technique is so involved.

Classes are in full swing, now. I had my first quiz in Cellular and Developmental Neuroscience this week, and both of my weekly seminars have started up, too. There are also lab meetings to attend, and meetings with my rotation adviser to plan. With all the other stuff going on I find myself working "late" (past 6 PM) several days a week, while a lot of my classmates seem to spend hardly any time at the lab. I guess it works out, though, since my lab time involves a lot of waiting around and doing other things. Even though I have to be physically present, I don't have to spend the entire day standing at my bench.