Membrane Type Contributing to High Background (Western Blot (Detection Problems))

Question

Accepted Answer

Persistent high background signal related to membrane choice, particularly when using PVDF membranes which generally produce more background than nitrocellulose. Common causes: 1) PVDF membrane inherently gives higher background than nitrocellulose membrane 2) Standard PVDF membrane unsuitable for fluorescent detection due to autofluorescence 3) Improper PVDF membrane preparation (not pre-soaked in methanol) Solutions: 1) Switch to nitrocellulose membrane which generally gives less background than PVDF 2) For fluorescent western blot, use low fluorescence PVDF membrane specifically designed for fluorescence detection 3) Ensure proper PVDF preparation: pre-soak in methanol then transfer buffer before use