![]() ![]() Fish skin gelatin is also cheap however it should be noted that it isn’t appropriate for use with biotin labelled antibodies due to it containing biotin. The main drawback of BSA usage is it’s relatively high cost compared to other blocking solutions.įish skin gelatin based blocking solutions offer an excellent alternative to other blocking solutions due to its lack of mammalian proteins which reduces the risk of antibody cross-reactivity with the blocking buffer. BSA is normally prepared as a 3% dilution in PBS-T or TBS-T. There can however be issues with contamination from native immunoglobulins which can cause issues with cross-reactivity. Alkaline phosphatase antibodies can also be inhibited by some preparations of milk alongside biotin labelled antibodies being interfered with by the milk preparation.īSA is also widely used as it is suitable for all detection systems as it does not contain biotin or phosphopeptides. Milk based blocking solutions should not be used for any experiments involving phosphorylation specific antibodies due to the presence of casein which is a phosphoprotein that can interfere with detection. Due to the rapid growth of microbial contamination milk based solutions should be made up fresh for every application. Some preparations contain sodium azide as a preservative although it should be noted that azide can inhibit horseradish peroxidase, the most commonly used secondary antibody conjugate in western blots. The most commonly used blocking solution is a solution of 5% non-fat dry milk in PBS-T which works well for the vast majority of applications at a inexpensive price. Comparison between commonly used blocking buffers in western blotting.
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