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when choosing a primary antibody select one that has been validated for use in western blotting supporting images should be available ensure that the antibody you select has been raised against an immunogen from the correct host select a primary antibody from a different host species than the host species of the cells or tissue used to prepare your lysate check that the antibody has been tested on a lysate that is likely to contain the target protein you can check this using tools such as the human protein atlas for an antibody targeting a phosphorylated protein ensure that the antibody has been tested on a lysate from cells that have been treated to stimulate phosphorylation check that the molecular weight of the band of interest in the images on the suppliers website matches the expected molecular weight of your protein of interest for example for p53 we would expect to see a band around 53 kilodaltons only in cell lines where p53 is expressed online databases such as