Transthyretin

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Electron Microscopy

Sample loading #1

1. Take out uranyl acetate (4%) from refrigerator.
2. Dilute uranyl acetate to 1%.
3. Support the discharged grid using the special tweezers. Use the edge of the grids.
4. Apply 10 μL of sample solution onto the grid, and incubate for 2 min.
5. Wick off the solution using a piece of the Whatman filter.
6. Apply 10 μL of filtered UA 1% onto the grid, and incubate for 2 min.
7. Wick off the solution with Whatman filter.
8. Air-dry the grid.
9. Place the grids into the gird box.

Sample loading #2

1. Take out uranyl acetate (4%) from refrigerator.
2. Dilute uranyl acetate to 1%.
3. Support the discharged grid using the special tweezers. Use the edge of the grids.
4. Apply 5 μL of sample solution onto a piece of parafilm, drop off the grid on it, and incubate for 2 to 5 min.
5. Wick off the solution using a piece of the Whatman filter.
6. Apply 5 μL of filtered UA 1% onto the parafilm, drop off the grid on it, and incubate for 2 to 5 min.
7. Wick off the solution with Whatman filter.
8. Apply 5 μL of filtered deionized water onto the parafilm, drop off the grid on it, and incubate for 2 to 5 min.
9. Wick off water using the Whatman filter.
10. Air-dry the grid.
11. Place the grids into the gird box.


ThT Fluorescence (Not very effective)

1. Place 200µL of sample solution in a 96 well plate.
2. Incubate the plate at 37°C with agitation.
3. Use a Synergy HT microplate reader to monitore the emission at 485/20nm.