Go west, young man

I’m back, after a break from the blogosphere, and the lab. I was home for an extended Dusshera-Diwali break. Plus, there wasn’t much happening in the lab over the last month anyway, barring the usual culturing and maintaining of cell stocks. By the way, HeLa has been a bit of a temperamental cell line. I had two sets die on me last week, and that shook me up a little. (My guide says you can never take it easy when it comes to cell culture. Even if you’ve been working with the same cell line for years, decades even, and you know all the ins and outs of your cell line, you can never let your guard down)

 

Anyway, I’m going to run my first western blot today. A western blot is an assay to check for the presence of a specific protein, first by separation based on size and then getting antibodies specific to that protein to bind with it. We’ll know that the antibody has bound to the target protein because it fluoresces on binding. I’m excited because:

a) It’s my first time

b) This is where my project really begins

 

If I haven’t already written about what I’m working on, here goes. My guide is a cancer biologist. She studies the proteins that are activated due to DNA damage. Some of these proteins arrest cell division, till the damage is repaired. It is suspected that these proteins don’t work the way they’re supposed to in cancer cells. We’re trying to confirm that that is what’s happening. Currently, I’m trying to induce DNA damage in HeLa and 293T and then see how expression of these proteins varies from a normal cell from either line. That is, of course, when my cells aren’t dying on me :(

 

I’ll be back later this week with a complete post on how my first western blot went. For now, I’m going to leave you with a bit of trivia. The reason western blotting is called western blotting is because it’s a play on Southern blotting (always a capital ‘S’). Southern blotting was developed by Edwin Southern in 1975 to check for specific DNA. When a technique to quantify protein was developed some time later, they called it western blotting. For RNA, it’s called northern blotting. No, there’s no such thing as an eastern blot, even if it seems unfair. Yes, I know, scientists are a very imaginative lot (rolls eyes)