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'' | |||
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 | ||
'''RECIPES''' | |||
<table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | <table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | ||
Buffer #1 | |||
<tr> | <tr> | ||
<th><tt> </tt></th> | <th><tt> </tt></th> | ||
| Line 52: | Line 64: | ||
Buffer #2 | |||
<table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | <table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | ||
| Line 63: | Line 76: | ||
<th><tt> 2 μL </tt></th> | <th><tt> 2 μL </tt></th> | ||
</tr> | </tr> | ||
<tr> | <tr> | ||
| Line 79: | Line 91: | ||
GO-Juice | |||
<table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | <table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | ||
<tr> | <tr> | ||
<th><tt> </tt></th> | <th><tt> </tt></th> | ||
<th><tt> 20 μL GO Juice </tt></th> | <th><tt> 20 μL GO-Juice </tt></th> | ||
</tr> | </tr> | ||
Latest revision as of 13:54, 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 |
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 |
