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Why do the bands of a pre-stained protein marker also appear in a Western blot?
The pre-stained protein marker is composed of recombinant proteins, and the protein markers used may contain a His tag. If the primary antibody carries a His tag, the secondary antibody will not only bind to the tagged primary antibody but also to the His‑tagged protein marker bands. Consequently, upon addition of the ECL luminescent substrate, both the protein marker and the target protein in the sample will be exposed simultaneously.
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