Today, I needed this hung on the wall next to my lab bench:
I’ll spare you all the moany, groany, whiney in-depth details, but suffice to say, my World’s Worst Protein is, once again, living up to it’s uncooperative title. I have spent the past 5 months trying to move forward with circular dichroism by dialyzing out the imidazole following elution (unsuccessful – always precipitates), dropping the pH (unsuccessful, still precipitates, and dropping it too low unfolds what has been refolded) and attempting to elute with EDTA. Finally (finally!) I was able to get the protein a) pure enough, b) refolded on the column, and c) off the column with EDTA, and today was the big reveal.
Suffice to say from the previous “Bang Head Here”, you can probably guess that rather than a chorus of angels descending from the heavens singing as a beautiful spectrum appeared as the result of all my hard work, thereby bringing me one step closer to maybe, possibly, potentially, someday graduating… I got an ugly bunch of sharp zig-zagging lines, because the amount of EDTA it takes to get my protein off the column apparently causes significant light scattering. Damn.
Nothing like watching 5 months of work unravel in 2 minutes. Bang head here, indeed.
Of course, while watching my spectra unfold in real-time, I happened to be Google Chatting with my best friend, which may have involved some explicit language and a pity-party for one regarding my inability to get just one decent looking spectra for Protein X. Within 3 seconds of the use of my word “spectra”, the followed magically appeared in my inbox from said best friend:
Yes, the “spectra” in reference is totally different, but the theory is the same. Learn more about the structure of the protein, and get a publication… and, while I can’t say it’s in my short-term plan like Calvin, I certainly wouldn’t turn down the Nobel Prize, the “get rich” money, or the talk show invitations. But Hobbes, I hate to break it to you – I don’t think scientists get too many babes.
P.S. Dear Protein, I hate you.
Oh Julie. That is so awful.
ReplyDeleteI wish you more luck in the very immediate future!
That stinks! Proteins, in general, are starting to get on my nerves. I hope that you can resolve the EDTA problem (more dialysis, I'm guessing) very soon!
ReplyDeleteYuck, sorry to hear about the shitty results. Knocking yourself unconscious will only be a temporary solution, I'm afraid. :)
ReplyDeleteUgh - sorry to hear about that! I deal with spectra too, and it can be very frustrating :S
ReplyDeleteI'm sorry for the terrible news.
ReplyDeleteI'm surprised more labs don't come with punching bags in the corner. A few rounds knocking out a protein or enzyme here or there would be very theraputic.
Maybe strangle a pipettman or two.
Boo! Stupid protein. Hopefully it'll learn its lesson sooner rather than later! Good luck!
ReplyDeleteStupid protein! I feel your pain, but my nemesis is actually a ~250 fragment of DNA. How can something so small be such a PITA? I don't know about you, but wine certainly helps me...
ReplyDeleteI am sorry to hear that Julie! :(:(:(
ReplyDeletethat sucks major ass :((
ReplyDeletebummer. :(
ReplyDeleteWe have that sign hanging up in our break room. I have been tempted to use it several times in the last 6 months.
what about using a different chelating agent to help knock it off the column.... you could check blanks of the chelators at high conc in the CD couldn't you?
ReplyDeleteargh, I'm sorry. But that comic is awsome.
ReplyDeletethat sucks bug time, sorry! On a funny note though, my next door bench mate has exactly the same Calvin & Hobbes cartoon posted on hir desk :)
ReplyDeleteI hate your protein too. And I don't even have to deal with it. Sorry to hear the latest protein update. It sucks alot.
ReplyDelete