Related Resources: Brochures | Application NotesBefore you start analyzing a protein sample by Western blotting, read our web page, “Protein Preparation,” for technical and product information on this important part of the Western blotting workflow. Often, the first step of analyzing protein expression or protein-protein interactions is to obtain a cell or tissue sample, lyse the cells, and extract proteins using extraction reagents. Total protein concentration must be determined for these cell lysates. Variables affecting each of these steps are outlined below, as each could affect the sensitivity and reproducibility of the Western blot.
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Different proteins may be best visualized via Western blot using different lysis buffers. Consider the pH, buffering compound, detergents and other additives when determining the optimal buffer for a specific protein.