Altman:Protocols/Single Molecule Assays/gliding filament
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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
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 |