So I am not sure exactly what happened but after adding my ECL reagent, part of the WB turned "brown". When imaging the blot it has a "burnt" effect as per below. It maybe that I had a high concentration of antibodies either the primary or the secondary. Is there anything I can do to save the WB?
Thank you :)
Adam B Shapiro
I don't think you can save the blot. It has been damaged from excessive peroxide formation.
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Reza Hadavi
hi
you can make one of the following changes :
1- Reduce exposure time to get the photo by camera
2- Reduce secondary antibody concentration (HRP-conjugated )
3- Reduce amount of cell lysate ( 10 or 15 ug/well loaded )
good luck
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