Altman:Protocols/Single Molecule Assays/single bead

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Revision as of 15:56, 15 June 2013


Department of Physics, Willamette University

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Single Bead Optical Trapping Assay

Before you start

  • Prepare an aliquot of α-GFP/10 (5 μL of α-GFP 0.5 mg/mL stock + 45 μL AB)

Make α-GFP beads


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

Buffer #1
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


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