User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2010/04/01/Osmotic stress planning: Difference between revisions

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{{AndyMaloneyNotebook
{{AndyMaloneyNotebook
  |Description = This notebook entry will be ultimately for planning experiments designed to look at kinesin and microtubules under osmotic stress. If I do run an experiment today, I will update the description to indicate that.
  |Description = This notebook entry plans for using betaine as an osmotic stessor and I note preliminary results.
}}
}}
[[Category:AM_OsmoticStress]]
[[Category:AM_OsmoticStress]]
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* Betaine will only be added to motility solutions. This means I only need 2M betaine in PEM. I will dub this solution PEM-Bine. (Ultimately I will look at sarcosine and proline and those solutions will be called PEM-Sine and PEM-Pine respectively.)
* Betaine will only be added to motility solutions. This means I only need 2M betaine in PEM. I will dub this solution PEM-Bine. (Ultimately I will look at sarcosine and proline and those solutions will be called PEM-Sine and PEM-Pine respectively.)
* Betaine is soluble to about 5 M in water. So, I'm going to make a 5 M stock solution of Betaine in PEM (PEM-Bine).
* Betaine is soluble to about 5 M in water. So, I'm going to make a 5 M stock solution of Betaine in PEM (PEM-Bine).
==Preliminary results==
So I ran the 1000mM betaine experiment today. Below are two movies. Both are of gliding motility assays with alpha casein as the surface passivation. The second movies is with 1000mM betaine in solution.
<html>
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I was really hoping for a dramatic affect but it doesn't look like there is one. Hmm...

Revision as of 16:30, 1 April 2010

Planning

The first chemical I am going to use as an osmotic stessor will be betaine. I want to complete 3 initial points to the data taking. 2 extreme points and one point between. I already have my 0 mM betaine point so I will take data for the following:

  • 1000 mM betaine
  • 5 mM betaine
  • 60 mM betaine

I understand that the 60 mM point isn't really between 1000 and 5 but, I am going to approach 1000 mM logarithmically. So if I approximately double the concentration from 5 to 1000, 60 is the middle point for me and what I plan to do.

Software needed

Since I've adopted the term software to mean chemicals, biological materials, and solutions, I need to prepare a 2M solution of betaine in PEM. There are some things to take into consideration when preparing this solution.

  • I don't want the betaine to directly interact with my casein. This means that no betaine will go into my casein stocks.
  • Due to my preliminary data on caseins, I will use alpha casein for my surface passivator.
  • Betaine will only be added to motility solutions. This means I only need 2M betaine in PEM. I will dub this solution PEM-Bine. (Ultimately I will look at sarcosine and proline and those solutions will be called PEM-Sine and PEM-Pine respectively.)
  • Betaine is soluble to about 5 M in water. So, I'm going to make a 5 M stock solution of Betaine in PEM (PEM-Bine).

Preliminary results

So I ran the 1000mM betaine experiment today. Below are two movies. Both are of gliding motility assays with alpha casein as the surface passivation. The second movies is with 1000mM betaine in solution.

<html> <object width="480" height="385"><param name="movie" value="http://www.youtube.com/v/JptG9hWi2ys&hl=en_US&fs=1&rel=0"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/JptG9hWi2ys&hl=en_US&fs=1&rel=0" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="480" height="385"></embed></object> </html>

<html> <object width="480" height="385"><param name="movie" value="http://www.youtube.com/v/ZAr773TKguU&hl=en_US&fs=1&rel=0"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/ZAr773TKguU&hl=en_US&fs=1&rel=0" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="480" height="385"></embed></object> </html>

I was really hoping for a dramatic affect but it doesn't look like there is one. Hmm...

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