Koch Lab:Protocols/Kinesin/Tubulin storage buffer

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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.

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