User:Nadiezda Fernandez-Oropeza/Notebook/Notebook/2011/05/18: Difference between revisions
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== | ==Gliding Motility Assay== | ||
* | * This entry describes the procedure in the Gliding Motility Assay | ||
===Procedure=== | |||
# First turn on the mercury lamp and prepare the microscope for Kohller Illumination. | |||
# Polymerize the microtubules (Procedure can be found here: [[User:Nadiezda_Fernandez-Oropeza/Notebook/Notebook/2011/05/16|Microtubule polymeration]]). | |||
# In the 30 minute wait for the polymerization of the microtubules, one can prepare the following solutions: | |||
#* 0.5 mg/mL of α-Casein in PEM: 500 µL of 1mg/mL α–Casein in PEM and 500 mL of PEM. | |||
#* PEM–αA: 1µL of PEM-A and 99 µL of 0.5 mg/mL of α-Casein in PEM. | |||
#* Kinesin in PEM-αA: 2 µL of Kinesin with 18 µL of PEM-αA. | |||
# In a flow cell, let 10 µL of 1mg/mL α–Casein in PEM rest for 10 minutes. | |||
# During this 10 minutes, prepare the Motility solution (Procedure can be found here: [[User:Nadiezda_Fernandez-Oropeza/Notebook/Notebook/2011/05/17#Motility_solution|Motility solution]]). | |||
# 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. | |||
# After those five minutes, exchange the fluid on the cell with 20 µL of the motility solution. | |||
# Finally, seal the end of the sample section on the flow cell with nail polish. | |||
Revision as of 18:38, 6 June 2011
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Gliding Motility Assay
Procedure
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