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. 2013 Jan 18;8(1):e54151. doi: 10.1371/journal.pone.0054151

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© 2013 Kaitaniemi et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Lysates of HEK293 cells co-transfected with flag-tagged VAP-1 cDNA, myc-tagged VAP-1Δ3 cDNA or with the corresponding tagged empty vectors in different combinations were separated in SDS-PAGE (with or without prior immunoprecipitation) and blotted to nitrocellulose membranes. The functionality of the tagged constructs and the ability to detect the proteins with corresponding antibodies was first verified by using the flag-antibody (A) or the myc-antibody (B) in control gels without prior immunoprecipitations. Aliquots of the same lysates were then immunoprecipitated with the flag antibody prior to gel electrophoresis, and the immunoprecipitated product was detected using the myc-antibody (C). The sizes of the two VAP-1 isoforms and the molecular weight markers are indicated. Ip Immunoprecipitation, Ig Immunoglobulin.