User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/11/05/Water isotope study
10% H-PEM 2nd try
So I'm not sure if it is because of the assay using D2O or if it is my alpha-PEM but, I'm not getting anything to stick.
Oh man. I'm going to go bananas if kinesin + alpha-casein + D2O have some odd interactions that are causing the kinesin to not adhere to the casein. Using different casein passivations cause differences in speed measurements and I've already done all of my O-PEM measurements using alpha casein.
I should point out that I am seeing some motility however, the motility is very similar to that of the regular assay using kappa casein. It took nearly 10 minutes for me to find 1 MT that was moving on the surface and it didn't move very well.
I'm going to try this again but this time I will make new H-PEM and see if that's the problem.
Nope. Same issues. I think I'll replace the alpha casein with whole casein.
This is interesting. I used the whole casein passivation and I'm motility. Really bad motility but I'm getting it. I'm going to see if it's the kinesin and use the alpha casein passivation.
Andy Maloney 13:28, 5 November 2010 (EDT): Phew! It was the kinesin. I'm again using the alpha casein passivation with new kinesin and it is working. Except of course for the first ROI where things are breaking up. Not sure why this happens but it may be due to the thermal shock the slide goes through when getting on the objective plus the fact that the MTs aren't moving around very much.
