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welcome to St Johnamp;#39;s laboratory video protocol Series today we will demonstrate to you how to carry out a western blot Western blotting allows you to detect and quantify specific proteins in a tissue sample the proteins in your sample must first be separated by size using gel electroforesis to prepare your protein sample combine the sample loading buffer the reducing agent distilled water and your protein sample in an endorf tube boil your sample for 10 minutes in a preheated heating block set to 80 C so that only the primary structure remains meanwhile prepare 800 ml of the running buffer take 200 mL from this and add 500 microliters of antioxidant this ensures that the protein maintains A reduced State having removed the gel from its packaging rinse it with distilled water and remove the plastic Combs to create the wells place it in your Western block gel tank and clamp it in place add the running buffer to the tank with the antioxidant in the inner compartme
