Dear All,
When I developed the protein signal with ECL on PVDF membrane, it displays many scratch marks background on the membrane. The blot was covered in plastic sheet to prevent drying. These scrath marks are the same one that appear when the membrane white-dried. (picture attached)
This membrane has been stripped two-three times before this time reprobling. The membrane was white-dried and resoak in 100%methanol for a few minute until the membrane turn grey, then washed with TBST before stripping>blocking with milk>reprobing. The primary antibody that was used in this time is high background one. So I washed it about 1hr wiht many change before developping.
On the previous stripping&reprobing with other antibodies, this problem had never occured, even the blot whited-dried and scratch marks appeared during the development.
What is a cause of the problem? How can I solve it? Can I sill use this membrane to probe this antibody?
Cheerios,
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