Difference between revisions of "Altman:Protocols/Single Molecule Assays/gliding filament"

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<th><tt> 100x PCr stock </tt></th>
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<th><tt> 20 μL GO Juice  </tt></th>
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<th><tt> 20 μL GO-Juice  </tt></th>
 
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Revision as of 12:53, 15 June 2013

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Department of Physics, Willamette University

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Gliding Filament Assay

Typical flow-through volumes are 10-15 μL

1. Make a flow cell with a nitrocellulose-coated coverslip

2. Flow in α-GFP/10

3. Incubate 2 minutes

4. Flow in ABSA

5. Incubate 2 minutes

6. Flow in motor dilution

7. Incubate 2 minutes

8. Flow in ABSA

9. Flow in Actin dilution

10. Incubate 2 minutes

11. Flow in GO-Juice


Recipes

Buffer #1
50 uL Buffer #1
50x GOC stock 2 μL
100x CPK stock 1 μL
100x ATP stock 1 μL
AB 46 μL

Buffer #2

50 μL Buffer #2
50x Glu/B stock 2 μL
100x PCr stock 1 μL
AB 47 μL


GO-Juice

20 μL GO-Juice
Buffer #1 10 μL
Buffer #2 10 μL