Hello friends,
We treated SH-SY5Y cells with 50 mM and 100 mM KCL to induce alpha synuclein accumulation. After 24 and 48 hours, we detected the aggregates of alpha synuclein by Zeiss microscope, but there was a staining over the nucleus. Below I will add some photos of the cells of interest.
My question is; why did the stain spread over the nucleus? Of note, we incubated primary antibodies overnight.
Hello Mohamed Khashan
In the paper attached, the investigators have suggested that nuclear staining by some of the alpha synuclein antibodies may result from their non-specific cross-reactivity to other antigens with epitope similar to that of alpha synuclein.
You may refer to the paper below for more information.
Article Determining nuclear localization of alpha-synuclein in mouse brains
Best.
As somebody who is in the field, i have to say most of the antibodies for α synuclein are not specific when it comes to IF. Some usually work with western blot, but we have had no luck with any antibody. Hence in pur studies we usually resort to fluorescent tagging of exogenous α synuclein, or transfect cells with α synuclein construct for endogenous α synuclein. Hope this helps.
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