Trichloroacetic Acid precipitation of proteins - (Jul/25/2014 )
I found this protocol for Trichloroacetic Acid precipitation of protein.
I am searching for the original paper by LUIS SANCHEZ, but i can't seem to find it.
Can anyone please help me? Send me a link or the paper here or at email@example.com
your link doesn't work and this should be in the "paper request" forum.
To TCA ppt proteins for dilute sds-page samples, this method has worked quite well for me:
1. pipette ~5-20ug of protein into a clean eppe tube
2. Add TCA (from a 100% (w/v) stock) to your protein solution to a final concentration of 20-30%
3. Let sit on ice for 15 min
4. Spin at 16k x g in a refrigerated centrifuge for 10 min
5. Aspirate off supernatant being sure not to disturb the tiny white pellet at the bottom
6. Wash pellet with ~1 ml cold (-20C) 100% acetone
7. Centrifuge again at 16k x g for 10 min
8. Aspirate off acetone being sure not to disturb tiny white pellet
9. Place empty tube with pellet in 95C heat block for 1 minute to dry off acetone (don't let it go longer than a minute)
10. Add 10-20ul of 1x SDS-PAGE sample buffer, vortex for a few minutes to fully resuspend pellet
11. Cook sample at 95C for 5 min
12. Load entire sample on gel.