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        TCA protein precipitation

        互联网

        1416

        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.

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