Altman:Protocols/Single Molecule Assays/gliding filament: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
David Altman (talk | contribs) No edit summary |
David Altman (talk | contribs) No edit summary |
||
| Line 34: | Line 34: | ||
<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 62: | Line 63: | ||
</table> | </table> | ||
Buffer #2 | |||
<table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | <table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | ||
| Line 88: | Line 90: | ||
GO-Juice | |||
<table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | <table id="Table 1" border="1" cellpadding="3" cellspacing="0"> | ||
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 |
