User:Nadiezda Fernandez-Oropeza/Notebook/Notebook/2011/05/18: Difference between revisions
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# First turn on the mercury lamp and prepare the microscope for Kohller Illumination. | # 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]]). | # 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: | # 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. | #* 0.5 mg/mL of α-Casein in PEM: 500 µL of 1mg/mL α–Casein in PEM and 500 mL of PEM. | ||
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#* Kinesin in PEM-αA: 2 µL of Kinesin with 18 µL of PEM-αA. | #* 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. | # 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]]). | # 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 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. | # After those five minutes, exchange the fluid on the cell with 20 µL of the motility solution. |
Revision as of 18:38, 6 June 2011
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Gliding Motility Assay
Procedure
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