Altman:Protocols/Single Molecule Assays/gliding filament

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==Making Nitrocellulose Coverslips==
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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
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2. Flow in α-GFP/10
2. Flow in α-GFP/10
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3. Incubate 2 minutes
3. Incubate 2 minutes
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4. Flow in ABSA
4. Flow in ABSA
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5. Incubate 2 minutes
5. Incubate 2 minutes
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6. Flow in motor dilution
6. Flow in motor dilution
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7. Incubate 2 minutes
7. Incubate 2 minutes
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8. Flow in ABSA
8. Flow in ABSA
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9. Flow in Actin dilution  
9. Flow in Actin dilution  
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10. Incubate 2 minutes
10. Incubate 2 minutes
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11. Flow in GO-Juice
11. Flow in GO-Juice

Revision as of 15:50, 15 June 2013


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

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


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