User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/07/29/Beta casein test: Difference between revisions
Andy Maloney (talk | contribs) |
Andy Maloney (talk | contribs) No edit summary |
||
Line 1: | Line 1: | ||
{{AndyMaloneyNotebook | {{AndyMaloneyNotebook | ||
|Description= | |Description=Trying the beta casein assay without putting it in the freezer and without filtering it and not putting it in the freezer. Guess what, it was the filtering. | ||
}} | }} | ||
[[Category:Kinesin and microtubules]] | [[Category:Kinesin and microtubules]] | ||
Line 23: | Line 23: | ||
After 1.5 hours, nothing. | After 1.5 hours, nothing. | ||
==Second try== | |||
I know I said I would stop but I did it again anyway. This time without filtering the beta casein solution. The assay works. | |||
Again, I find that I am overwhelmingly infuriated at myself for not writing this down earlier. All this trouble is because of my lack of notebook entries. |
Revision as of 13:29, 29 July 2010
Yesterday, I found out that none of my casein solutions were supporting motility except for my whole casein. I have narrowed the reasons for this down to the possibility that it was storing them in the -80C freezer. Today, I prepared more beta-PEM and I'm going to try an assay to see if it works. I did send the solution through a 0.2um filter.
First try
Nope...No motility on the slip. I did see a small amount on the slide but I can't use that. I'll wait a few more minutes to see if anything is working.
15 minutes in and still nothing.
I don't get it. It just isn't working. And of course I didn't save some beta-PEM to not filter it so I could see if that was the issue.
- The Taxol is working just fine because the MTs are stable.
- The Antifade is working because after blasting them with 100% illumination, they are still bright.
- The kinesin is good as has been shown with the whole casein assay. As well as the PEM-Glu and PEM-A are good because whole casein works.
- The freezer isn't the problem as was shown right now.
- The filtering can't be the problem because the whole casein works. Unless the filter is gobbling up all the casein. But then I wouldn't see bubbles forming in my casein solutions, which I do so there is protein in them.
This truly has become a mystery now. Maybe if I get a new batch of kinesin I'll try this again. But, I'm out of ideas and there is no point trying to brute force anything now.
After 1.5 hours, nothing.
Second try
I know I said I would stop but I did it again anyway. This time without filtering the beta casein solution. The assay works.
Again, I find that I am overwhelmingly infuriated at myself for not writing this down earlier. All this trouble is because of my lack of notebook entries.