New Resource: Western blotting (WB) Guide and Troubleshooting

Western blotting is a laboratory technique used by life science researchers and diagnostic laboratories to detect specific proteins within a homogenate or extract of a biological sample.  Before immunological detection, sample proteins must be extracted from their source using an optimized lysis buffer, separated within a gel using an electrophoresis unit and transferred to a membrane via a “blotting” procedure.  Proteins can be separated within a gel by the size of their individual polypeptide subunits to determine their molecular weight, or they can be separated based on their native, tertiary form such that subunit interaction and conformation are preserved.  The latter form of separation is determined by their charge-to-mass ratio.  After the proteins have been immobilized on the membrane, the proteins of interest can be identified through immunological detection using antibodies conjugated for visualization via chromogenic, chemiluminescent, fluorescent or radioactive mechanisms.

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