Validation of phospho-eEF2 protein as the target of autoantibodies in mice bearing colorectal HCT116 tumors.

Abstract

<p><b>A.</b> Representative immunoblotting of 2D-separated lysates of hypoxic HCT116 cells with a commercial antibody against eEF2. Proteins of the lysates are labelled with Cy dye (red) and secondary antibody is conjugated to horseradish peroxidase (green spots). Positive signal is obtained for several spots of the same molecular weight but differing by their pI value. <b>B.</b> Comparison of the eEF2 spots detected with a commercial antibody against total eEF2 (top), the serum from tumor-bearing mice (middle) and a commercial antibody against phospho-Thr56 eEF2 (bottom). Spot 4 (rightmost spot) corresponds to the unphosphorylated form of eEF2 while the other spots correspond to multi-phosphorylated forms of the protein; spot 3 (second spot from the right) corresponds to the preferential monophosphorylated form of eEF2 (on Thr56).</p

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