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Unspecific Western Signal from Protein A tag?

Western Blotting

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#1 michael_p



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Posted 29 January 2015 - 03:13 AM



I`m running a western blot of lysates from cells overexpressing proteins with a ZZ-domain (part of protein A). I observe non-specific signal from my GAPDH-antibody from the same locations where I detect the protein (this is a fluorescence based western with incubation with both antibodies simultaneously). I was wondering if it is possible that the protein A tag will -despite denaturation and reduction- still bind any antibody. (My GAPDH antibody is from mouse). If this is the case does anyone have a suggestion how to prevent this?

Thank you!

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