The proteins serving as loading controls must fit certain criteria. The number of publications (num) that cites each type of main loading controls with antibodies from each of the major suppliers. Other interesting observations from this survey are discussed in this article later. The results indicate that actin (specifically beta actin) is the most commonly used control. Figure 1 displays the Western blot images compiled from some of those publications. Labome has surveyed publications with loading controls in Western blots (Table 1). Looking to the literature can be very useful in considering potential loading controls for your Western blotting experiments. For this reason housekeeping genes that are constitutively expressed and necessary for basic cell functions, like cytoskeletal proteins, are typically used for loading controls. For any particular experiment it is important to consider potential loading controls and try to find a protein that is expressed at stable levels among the samples of interest. A variety of different proteins are used as loading controls. To use a loading control for these purposes detection with control protein antibody and the experimental antibody should be done on the same blot. The signals from loading controls are typically used to normalize the signals from the proteins of interest. - antibody incubation (for primary or secondary antibody), and signal detection across separate samples/lanes.- protein transfer efficiencies from the gel to the membrane among different samples/lanes,.- the amount of protein sample loaded in each lane,.Ī loading control accounts for potential variation in:
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