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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