Altman:Protocols/Single Molecule Assays/single bead

Department of Physics, Willamette University

'

Home        Contact        Lab Members        Publications        Research        Protocols        Pictures       

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