![]() For this reason, ‘housekeeping’ genes are frequently chosen as loading controls. What makes a good loading control you ask? You should look for proteins that exhibit high level, constitutive expression in the cell type you are examining, as this means it will have constant high expression levels. They can be used to quantify the protein amounts in each lane and normalise the expression levels of your protein of interest to the loading control Once you have established there are not large differences in protein levels, loading controls can then be used to normalise small variations in levels.Loading controls can show if this has occurred and allows for this variation to be accounted for. This can result in more intense stains at the edge than in the middle of the gel. This helps to protect against what is known as the “edge effect”, that is when large numbers of lanes are being used, proteins in the outer lanes transfer to the membrane in a position close to the frame.For example, they can be used to check that there has been even transfer from the gel to the membrane across the whole gel.They allow you to check that protein loading is the same across the gel, the expression level of the loading control should not vary between the different sample lanes. Loading controls are essential for publication worthy Western Blots.Why do I need a loading control for Western blotting? We provide a guide to some of the most commonly used types of loading control antibodies. Antibodies against several different targets are commonly used and it’s not always obvious which you should choose. Loading control antibodies are some of the most utilised antibodies in research.
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