Template:SBB- Surface-binding Assay

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All incubation steps are done in a normal incubator with no shaking.

1) Prepare a solution of your binding target (gliadin, etc.) at an appropriate concentration in a buffer that the target is soluble in.

2) Add 100 μL of the target solution per well to a 96-well MaxiSorp plate.

3) Incubate at 37C for 1 hr.

4) Plate wash 3x with 100 μL PBS.

5) Add 100 uL of 1.0 mg/mL BSA in PBS.

6) Incubate at 37C for 30 min.

7) Aspirate the BSA solution, then add the bacteria (about 5uL bacteria + 95uL of 1.0 mg/mL BSA in PBS).

8) Incubate at 37degC for 40 min

9) Plate wash 3x with 100 μL PBS. Uniformity is key, so use the plate washer at its gentlest settings.

10) Add 100uL of LB with appropriate antibiotics.

11) Cover with clear tape and put in the incubator.

12) Watch them between 3 hr and 5 hr of incubation for cloudiness and note how long it took to appear. Alternatively: TECAN the plate for OD600 at 3, 3.5, 4, 4.5, and 5 hours.