What is the difference between a Western blot performed under reducing versus non-reducing conditions?
To perform a Western blot under reducing conditions simply include a reducing agent such as dithiothreitol (DTT) or β-mercaptoethanol in the SDS sample buffer. These reducing agents act to disrupt any disulfide bonds. The 150 kDa IgG molecule, for example, will be separated into 2x heavy (~50 kDa) and 2x light chains (~25 kDa).
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