Western blotting is an extensively used technique for protein analysis. After transfer, mark the orientation of the gel on the membrane.The western blot is a common method used to detect proteins as well as post-translational modifications on proteins, and can provide semi-quantitative or quantitative data about the target protein in simple or complex biological samples.The field strength required is determined by the surface area and thickness of the gel: 0.8 mA/cm2 is a useful guide (1 h transfer). Transfer efficiency should be monitored by staining (see below). Tip: Time of transfer is dependent on the size of the proteins, percentage acrylamide, and gel thickness. For current, voltage, and transfer times specific to your apparatus, consult the manufacturer’s instructions. ![]() They can be removed by gently rolling a Pasteur pipet over each layer in the sandwich. Tip: Air bubbles may cause localized nontransfer of proteins.
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