Hijinks and Triumphs
Currently in my project I have to do a large series of experiments called Western Blots. Without going into too much detail, Westerns can be a little laborious (in that they have a lot of steps and can take a long time.) They are involve techniques that are relatively standard unless you are me. I almost never work with proteins, so the initial steps of running a protein gel and then transferring the gel to a membrane seem slightly like brain surgery as far as I am concerned. Until last week, I had done 1 Western in my life and it was 6 years ago and I had never run a protein gel or done any other protein work outside of that one long-forgotten Western.So, last week JP the wonderful tech and I started on my massive series of Westerns. The whole process was extremely painful and involved lots of crazy problems and hangups. Unsurprisingly, the experiment failed. When JP was asked why, she said "It was sketchy." When I was asked why, I said "Too many hijinks."
Today I am repeating the Western. Unfortunately, JP is busy with other work, so I'm on my own. This is not good, especially since I was rather distracted during the beginning steps last week...I was answering other peoples' questions and trying to do lots of things and sort of missed our lesson on how to set everything up. However, the experiment is underway! I have managed to set up the first major step (running the gel) with rather minimal help. Only one difficult step left.
The good news about Westerns? They involve a lot of incubation steps (as in set up gel, wait 90 minutes. Set up transfer, wait 2 hours. Etc.) which means that once I get through the pain of setup (that will become easier with practice) I have play time. :)
posted by Immunegirl @ 1:46 PM
9 Comments:
Western omelette mmmmmmmmmmm :-)
So? How did it go? Just don't blow yourself up.
It's going okay so far. I'm currently in the middle of the two hour transfer. I needed a bit of help to set up the transfer, but it seems to be going okay.
Good. While you are waiting for the results, can you make me a western omelette? *sheepish grin*
One Western Omlette coming up. Do you mind if it has some of this lab buffer in it? It's really just the same detergent that is in shampoo....
So, you're telling me Mr Bob spends all his time slacking off when he complains about Western blots?
No....they really suck to set up. But they DO allow free time. :)
A little buffer never hurt anyone.
Thanks Iggy!
Dood. I never did a Western, but I did so many protein gels I could probably still do them in my sleep, even after six years away (holy crap! I am old).
Here's a helpful hint: don't drink the acrylamide.
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