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TCA Precipitation with DOC

TCA Precipitation with DOC

 

Reagents:

2% deoxycholate DOC

24% trichloroacetic acid (TCA)

ice cold acetone

1X PAGE sample buffer

Protocol:

1. Add 2% DOC to 0.02% final (for 100 µl sample, add 1 µl 2% DOC).

2. Incubate at room temperature for 15 min.

3. Add 24% TCA to 6% final (for 100 µl sample, add 33.7 µl).

4. Incubate on ice for 1 hour.

5. Microfuge at 4 oC for 10 min, remove supernatant with drawn-out Pasteur pipette.

6. Add 200 µl of ice cold acetone to TCA pellet.

7. Incubate for 15 min on ice.

8. Microfuge at 4 oC for 10 min.

9. Remove supernatant as before, dry pellet for 1-2 min.

10. Resuspend pellet in 1X PAGE sample buffer ---> to gel.

 

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revised 6/24/1998 cla