User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2009/10/30/Heavy motility

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Heavy PEM

So I have polymerized new microtubules and I have fixed them with a solution of Heavy PEM-T. I also have solutions of

  • Heavy PEM-[math]\displaystyle{ \kappa }[/math]C
  • Heavy PEM-[math]\displaystyle{ \kappa }[/math]CT
  • Heavy PEM-[math]\displaystyle{ \kappa }[/math]CA

This means I can start a motility assay with the heavy water stuff.

Last time I tried this I was able to polymerize and stabilize microtubules in a solution of Heavy PEM. Hopefully, this will work this time around as well.

"100%" Heavy PEM

So 100% is in quotations because it really isn't 100% heavy water. I did not store my microtubules in a solution of heavy water nor is my kinesin in a solution of heavy water. However, every other buffer does have heavy water in it and my PEM is made from 100% heavy water.

As you can see, the results speak for themselves. Not only are microtubules stable in Heavy PEM, but the kinesin does process!

<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/gXStPSVnDNY&hl=en&fs=1&"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/gXStPSVnDNY&hl=en&fs=1&" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="425" height="344"></embed></object> </html>

After 30 minutes, the little guys are still going!

<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/R_MsiMUko8c&hl=en&fs=1&"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/R_MsiMUko8c&hl=en&fs=1&" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="425" height="344"></embed></object> </html>

And 1 hour and 30 minutes later.

<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/ueHYxSkxkOs&hl=en&fs=1&"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/ueHYxSkxkOs&hl=en&fs=1&" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="425" height="344"></embed></object> </html>

Okay, there are just way too many videos to upload right now of really cool stuff. I will have to do it tomorrow.

Steve Koch 00:24, 31 October 2009 (EDT): My head is still spinning from all the cool things you saw with that assay. Too many thoughts to write down, but have to say, CONGRATULATIONS! I think this is a huge breakthrough. Even without knowing much fundamental, it's fascinating. But I think it also shows that we have a new, crucial knob that we can turn in this system. I can't figure out what category this page should be. "Instant Classic" "Breakthrough" "Awesome"
Steve Koch 00:25, 31 October 2009 (EDT): Also, I am confused as to how much to advertise this. I'm so excited, I feel like posting all over the place. But also wondering whether you should be the one to do that? Or whether you'd want to wait. A good connundrum to have. Let me know what you think :)
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