User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2009/08/27/Tubulin resuspension

Resuspension how to

Fill in later.

First attempt

I got a new vial of kinesin from the freezer and added that to a passivated glass surface which was passivated with [math]\displaystyle{ \kappa }[/math]-casein. We polymerized our newly resuspended tubulin which consisted of 15% labeled rhodamine tubulin and 85% unlabeled tubulin. While the polymerization worked beautifully, we forgot to add dextrose to the anti-fade and thus lost the microtubule sample quickly. Thankfully we were able to see the microtubules in solution very well with 15% rhodamine.

Another blow below the belt is that the first slide didn't have active kinesin. Well, maybe it was bad ATP and only another try will tell.

Steve Koch 23:27, 27 August 2009 (EDT): Not sure exactly what kicked you in the nuts, but most likely, I would say the lack of dextrose. It turns out that antifade not only helps with anti-photobleaching, but also is important for overall motility (if I remember correctly).

Andy Maloney 01:45, 28 August 2009 (EDT): I so don't understand that at all. Apparently though, it's true because the other blokes got it to work once they added the dextrose to the assay.