Koch Lab:Protocols/Kinesin/Tubulin storage buffer

Using previous protocols from Koch and Cytoskeleton, we have been making a tubulin storage/polymerization buffer that has been comprised of PEM with 6% glycerol (v/v) and 1 mM GTP.

You can purchase a tubulin storage buffer from Cytoskeleton which has 60% (v/v) glycerol in PEM. The only problem with this is that we have determined that we want tubulin to be in PEM that has a total concentration of 2 mM MgCl₂ because of having the chelator EGTA in solution as well. Because of this, we make our own tubulin storage/polymerization buffer.

Recipe

• PEM
• 6% (v/v) glycerol
• Additional 1 mM MgCl₂
• 1 mM GTP

Pipetting glycerol is not the easiest thing to do. Just be careful and mindful of how viscous it is.