User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2009/08/27/Tubulin resuspension: Difference between revisions
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[[User:Steven J. Koch|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). | [[User:Steven J. Koch|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). | ||
[[User:Andy Maloney|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. | |||
Revision as of 22:45, 27 August 2009
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.
