Koch Lab:Protocols/Kinesin

The Kinesin Home Page is an excellent resource. Many of our protocols probably evolved from there, which in turn were obtained from prior work of others. It's tough to know who to cite on these protocols, so please send us a note if you'd like us to cite it properly.

Chemicals

• ATP suspension
• GTP suspension
• Taxol suspension
• Glycerol
• [math]\displaystyle{ \kappa }[/math]-Casein

Buffers

• PEM
  • Koch's version of PEM

Steve Koch: I can't remember whether I made this buffer for use with malachite green inorganic phosphate assays, or because I thought it was better to use potassium ions versus sodium ions in kinesin assays.

• Tubulin storage buffer

Used to store as well as polymerize tubulin.

• PEM-T

Used to stabilize microtubules after polymerization.

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

Used to passivate glass in order to keep kinesin viable.

Methods

• Tubulin resuspension and polymerization (Cytoskeleton)
• AMP-PNP Purification of active kinesin
• Dynamic Light Scattering measurement of kinesin aggregation