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Do the bands of the pre-stained protein marker fade on the membrane during transfer?
Fading is most likely caused by detergents in the blocking or washing solutions, which strip some proteins from the membrane. The dyes themselves do not elute from the proteins because they are covalently bound. Membranes with smaller pore sizes have been shown to better retain proteins during blocking and washing; therefore, for optimal resolution and protein retention, a 0.2 µm membrane is recommended over a 0.45 µm membrane. After transfer, you can use a pencil to outline the pre-stained bands on the membrane, allowing you to identify band positions even after blocking and processing.
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