Altman:Protocols/Single Molecule Assays/gliding filament: Difference between revisions
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== | ==Gliding Filament Assay== | ||
''Typical flow-through volumes are 10-15 μL'' | ''Typical flow-through volumes are 10-15 μL'' | ||
1. Make a flow cell with a nitrocellulose-coated coverslip | 1. Make a flow cell with a nitrocellulose-coated coverslip | ||
2. Flow in α-GFP/10 | 2. Flow in α-GFP/10 | ||
3. Incubate 2 minutes | 3. Incubate 2 minutes | ||
4. Flow in ABSA | 4. Flow in ABSA | ||
5. Incubate 2 minutes | 5. Incubate 2 minutes | ||
6. Flow in motor dilution | 6. Flow in motor dilution | ||
7. Incubate 2 minutes | 7. Incubate 2 minutes | ||
8. Flow in ABSA | 8. Flow in ABSA | ||
9. Flow in Actin dilution | 9. Flow in Actin dilution | ||
10. Incubate 2 minutes | 10. Incubate 2 minutes | ||
11. Flow in GO-Juice | 11. Flow in GO-Juice | ||
Revision as of 13:50, 15 June 2013
Department of Physics, Willamette University
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
| 50 uL Buffer #1 | |
|---|---|
| 50x GOC stock | 2 μL |
| 100x CPK stock | 1 μL |
| 100x ATP stock | 1 μL |
| AB | 46 μL |
| 50 μL Buffer #2 | |
|---|---|
| 50x Glu/B stock | 2 μL |
| 100x PCr stock | 1 μL |
| AB | 47 μL |
| 20 μL GO Juice | |
|---|---|
| Buffer #1 | 10 μL |
| Buffer #2 | 10 μL |
