Dark room frisson
This is why I love science, love it so much that I couldn’t leave it for long.
Last night, late for my date and fretting in the dark room, the hot breath of the x-omat developer wafting over the stink of chemicals, I had absolutely no hope: I was trying to make an antibody work that everyone else had deemed a dud. I’d already tried a number of conditions to no avail, and this last one was just a punt because I’d had an extra strip that I hadn’t yet tested and I thought it was a shame to throw it away. So I bumped up the concentration and tried a different temperature, but feeding the film into the machine was just a formality.
Damned if the blot didn’t sport the most beautiful band I’ve seen for the past year, clean as the proverbial whistle.
Result.
This is exactly what I *don’t* like about bench work: you worked SO HARD with no result whatsoever, and then a throwaway experiment gives results. I want effort and result to be at least a positive correlation. Doesn’t have to be exponential, or even linear – just positive. With lab work it always felt like there was a negative correlation between the two.
Isn’t that the kind of tip that researchers should be willing to share with each other as practically a matter of routine – in some kind of antibody forum/database, say? Or maybe if you’d seen a bunch of people saying that they couldn’t get the antibody to work then you wouldn’t have even bothered to try yourself.
Yes, and Jenny’s talked about this very issue on a number of occasions. You’d think that Web 2.0 would be ideal for this sort of thing, but we have so many ‘Facebook for Science’s that the effort is too disperse to make it useful.
One for f1000 perhaps *cough*
Except that we made the antibody ourselves. So no one else in the world would know.
Even if it was commercial, it could vary. My entire PhD hinged on a particular vial of an antibody that was much stronger than the previous batch of the same antibody from the same supplier. And an epiphany, when I realized, in the shower, that I could use that extra strong batch to retry a previous crucial experiment in which I had such tiny numbers of cells that the bands on the blot were too faint to draw any conclusions. It worked, but it also made me realize how flaky biology is. Yes, it works *now*, but there’s no guarantee that you can repeat it if the commercially ordered product isn’t exactly the same every time.
Thing is, the _principle_ of changing temperature is what should be known. It’d be handy to have a list of conditions for all antibodies, but even handier to know that this is a trick to try.
Cool! It is worth the slog when you do get such a nice result though.