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MOLECULAR BIOLOGY: WORKING WITH DNA

PURIFICATION: PRECIPITATION

Trichloroacetic Acid Protein Precipitation

Trichloroacetic Acid Protein Precipitation Title | Overview | Procedure | Solutions | BioChemicals | Hints | Printable Version
Contributor:	The Laboratory of Steve Hahn at the Fred Hutchinson Cancer Research Center
Overview Title | Overview | Procedure | Solutions | BioChemicals | Hints | Printable Version
This protocol is used to concentrate proteins for further analysis (e.g., by sodium dodecyl sulfate polyacrylamide gel electophoresis [SDS PAGE]).
Procedure Title | Overview | Procedure | Solutions | BioChemicals | Hints | Printable Version
1. Add an equal volume of 20% TCA to the protein sample. 2. Incubate the sample for 30 min on ice. 3. Centrifuge the sample in a microcentrifuge at full speed for 15 min at 4°C. 4. Carefully remove all of the supernatant. 5. Add 300 μl of cold Acetone to the protein pellet and centrifuge again at full speed for 5 min at 4°C. 6. Remove the supernatant and allow the protein pellet to dry. 7. The protein can now be analyzed through SDS PAGE (see Protocol ID#455) by resuspendng the protein pellet in Sample Buffer. Heat the sample to 65°C for 3 min and load the protein onto a SDS polyacrylamide gel.
Solutions Title | Overview | Procedure | Solutions | BioChemicals | Hints | Printable Version
Sample Buffer    0.0025% (w/v) Bromophenol Blue 5% (v/v) 2-Mercaptoethanol 2% (w/v) SDS 10% (v/v) Glycerol 0.06 M Tris-HCl, pH 6.8 20% (w/v) Trichloroacetic Acid
BioReagents and Chemicals Title | Overview | Procedure | Solutions | BioChemicals | Hints | Printable Version
Glycerol 2-Mercaptoethanol Tris-HCl Trichloroacetic Acid Bromophenol Blue Sodium Dodecyl Sulfate Acetone
Protocol Hints Title | Overview | Procedure | Solutions | BioChemicals | Hints | Printable Version
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