User:Linh N Le/Notebook/2009/06/23: Difference between revisions
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* Try Kinesin? | * Try Kinesin? | ||
* Clean up lab | * Clean up lab | ||
12-6 | |||
==BRB80== | ==BRB80== | ||
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*What we use for Cytoskeleton is called PEM and while PEM and BRB80 ''can'' be the same, they do not ''have'' to be | *What we use for Cytoskeleton is called PEM and while PEM and BRB80 ''can'' be the same, they do not ''have'' to be | ||
**I did searches on both PEM and BRB80 and the following recipes are also reported by what they are called | **I did searches on both PEM and BRB80 and the following recipes are also reported by what they are called | ||
*''Special Note: All PDF's linked here are from the personal website of the authors and are free access'' | |||
===Recipes=== | ===Recipes=== | ||
*[http://www.proweb.org/kinesin/Methods/motility.html Hancock (Used by us as well)] | *[http://www.proweb.org/kinesin/Methods/motility.html Hancock (Used by us as well)] | ||
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***1 mM MgCl2 | ***1 mM MgCl2 | ||
***pH 6.9 | ***pH 6.9 | ||
*[http://www.stanford.edu/group/blocklab/Block%20et%20al%20PNAS%202003.pdf Mathew Lang, Steve Block first author] | |||
*Also used in [http://www.stanford.edu/group/blocklab/Rosenfeld%202003.pdf Rosenfield, Block Paper] | |||
*Also used in [http://www.stanford.edu/group/blocklab/Guydosh%20and%20Block.pdf Guydosh Block Paper] with 10μM of Taxol | |||
**Un-named buffer | |||
***80 mM Pipes (pH6.9) | |||
***50 mM potassium acetate | |||
***4 mM MgCl2 | |||
***2 mM DTT | |||
***1 mM EGTA | |||
***7 mMTaxol | |||
**Andy mentions that he thinks these buffers are what Block used to get kinesin to stick to beads. It not be what we want, but if the whole "soup" (Beads + kinesin + buffer) is added to the microtubes mix and it doesnt mess things up, this stuff should be fine. | |||
*[http://www.stanford.edu/group/blocklab/Valentine%20Fordyce%20et%20al%20NCB%202006.pdf Valentine, Fordyce, Block] | |||
**polymerization buffer for brain tubulin | |||
***62 mM PIPES at pH 6.9 | |||
***0.8 mM EGTA | |||
***3.7 mM MgCl2 | |||
***0.9 mM GTP | |||
***10% v/v DMSO | |||
**stablilzation buffer | |||
***80 mM PIPES at pH 6.9 | |||
***1.7 mM EGTA | |||
***5.5 mM MgCl2 | |||
***1.2 mM GTP | |||
***8.2 mM sodium azide | |||
***20 μM taxol | |||
***10% v/v DMSO) | |||
**Assay buffer based on PEM80 | |||
***80 mM PIPES at pH 6.9 | |||
***1 mM EGTA | |||
***4 mM MgCl2 | |||
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Revision as of 16:30, 24 June 2009
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To Do
12-6 BRB80
Recipes
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