User:Nadiezda Fernandez-Oropeza/Notebook/Notebook/2011/05/18

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Gliding Motility Assay

 * This entry describes the procedure in the Gliding Motility Assay

Procedure

 * 1) First turn on the mercury lamp and prepare the microscope for Kohller Illumination.
 * 2) Polymerize the microtubules (Procedure can be found here: Microtubule polymeration).
 * 3) In the 30 minute wait for the polymerization of the microtubules, one can prepare the following solutions:
 * 4) * 0.5 mg/mL of α-Casein in PEM: 500 µL of 1mg/mL α–Casein in PEM and 500 mL of PEM.
 * 5) * PEM–αA: 1µL of PEM-A and 99 µL of 0.5 mg/mL of α-Casein in PEM.
 * 6) * Kinesin in PEM-αA: 2 µL of Kinesin with 18 µL of PEM-αA.
 * 7) In a flow cell, let 10 µL of 1mg/mL α–Casein in PEM rest for 10 minutes.
 * 8) During this 10 minutes, prepare the Motility solution (Procedure can be found here:  Motility solution).
 * 9) After the 10 minutes are up, exchange the fluid on the cell with the solution of Kinesin in PEM-αA, and let it incubate for 5 minutes.
 * 10) After those five minutes, exchange the fluid on the cell with 20 µL of the motility solution.
 * 11) Finally, seal the end of the sample section on the flow cell with nail polish.


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