TCA protein precipitation
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Kyle Coachman/Linda Warfield/Steve Hahn
Last modified 5/13/02
To concentrate proteins for analysis by SDS PAGE:
If a small amount of protein is to be precipitated (less than a few micrograms),add Insulin as a carrier protein (10 micrograms of Sigma insulin,I-5500,per sample works well).
1.Add an equal volume of 20% TCA (trichloroacetic acid)to protein sample.
2.Incubate 30 min on ice.
3.Spin in microfuge at 4 deg.For 15 min.
4.Carefully remove all supernatant.
5.Add ~300μl cold acetone and spin 5 min at 4 degrees.
6.Remove supernatant and dry pellet.
7.Resuspend samples in SDS PAGE loading buffer.Load to SDS PAGE after heating at 65 deg for 3 min.
