Hoefer SE300 miniVE - Manual (Page 33)

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p25
Inefficient
binding Chemical
parameters Fix or crosslink the mole to the requirements of the nucleic acid,
protein, or membrane type.
Prepare protein transfer buffer without SDS. SDS can improve
transfer efficiency but reduces binding.
Verify the optimal amount of methanol required for the membrane
type and check the buffer solution. Add 10–20% methanol to the
transfer buffer to enhance binding to nitrocellulose.
Membrane
parameters Wear gloves when handling membranes.
Store membranes properly. Protect them from temperature
extremes and direct sunlight.
If proteins pass through the selected membrane, try a different
type or one with a smaller pore size (0.10–0.20 µm).
If different proteins may be migrating in opposite directions,
place a membrane on both sides of the gel.
If the sample load may be exceeding the capacity of the binding
surface area, apply two membranes. If “blow through” occurs,
reduce the sample load.
problem possible cause solution