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hello everyone this video covering the blocking step of a western blot will go over blocking reagents blocking techniques and the relevance and necessity of this step to start off what is blocking blocking prevents non-specific binding of antibody to the blotting membrane because the membranes typically used for a western blot have a high affinity for all proteins the primary and secondary antibody may bind not specifically to the membrane the result is what is referred to as high background or noisy blots here you can see an image of high background compared to ideal results blocking covers up these non-specific protein binding sites ensuring that most of your primary antibody binds to the target protein three to five percent non-fat milk or bsa are common reagents used these are diluted in tbs or pbs buffers finally how do you block during a western blot you will need tweezers your membrane a container your blocking reagent in this case non-fat milk and a shaker first you wil
