After the transfection, I blotted the membranes with commonly used beta-actin ab (sigma) in our lab. I saw 2 bands for the fırst time and I do not know the reason.
Thank you for your concern.
Sinem
What is the size of the second protein (band)? If it is larger, it is an aggregate or dimer. If it is smaller, then it is a cleaved product. Do you have negative controls? If it is present in negative control, then this is non-specific from cell extract.
There are many enzymes that can degrade actin. After degradation, typical fragments such as 35-38 kDa, ~30 kDa, and 15 kDa will be produced, among which the 35-38 kDa fragments are the most common.
Solution:
Add less primary antibody, shorter substrate reaction time, shorter exposure time
It might be due to the antibody
I would he recommend you check antibody on different protein ısolated fromcell line.
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