User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/08/04/Solutions
New solutions
So after figuring out that filtering the casein solutions through a 0.2um filter causes the solution to have not enough casein, I'm finally remaking them. This of course means making more PEM. In order to not have the issue of not enough casein blocking the surface, I am making new aliquots of
- alpha-PEM
- beta-PEM
- kappa-PEM
- whole-PEM
at the concentration of 1mg/mL for each casein constituent. Now, I recall that Verma et. al. used 0.2mg/mL casein to passivate the surface. I have typically been doing 0.5mg/mL passivation and then adding 0.25mg/mL casein in with my kinesin. Previously, before I filtered my casein solutions, this worked just fine. But, I'm now curious to see what happens when I use 1mg/mL to passivate the surface. This is why I have prepared solutions using 1mg/mL casein in them. Of course, I'm going to start off my experiments using the usual 0.5mg/mL passivation and then the 0.25mg/mL in the kinesin solutions. But, I think that all the troubles I was having with the kappa casein passivation may change if I use more protein to passivate the surface.
I should reiterate that the reason why I'm doing these casein studies is to see if there are any speed differences of gliding with temperature stabilization.
Andy Maloney 17:15, 4 August 2010 (EDT): Update
- I have finally almost completely finished making the new solutions. It appears that it takes about 8 hours to make everything properly. I will start new experiments tomorrow.