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. 2010 Apr 12;5(4):e10118. doi: 10.1371/journal.pone.0010118

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Zanetti 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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A) SynK and the monoclonal antibody 3A8 against TPK3/5 recognize the same, 54 kDa band in Arabidopsis wild-type thylakoids (proteins corresponding to 30 µg chorophyll were loaded). B) Intensity of the 54 kDa band decreased when the antibody was preincubated with 300 µM immunogenic peptide. The two lanes (30 µg Chl/lane) are from the same blot and were processed together. C) Thylakoids isolated from WT Arabidopsis plants were immunoprecipitated with anti-SynK antibody and blotted with 3A8 monoclonal antibody. D) Thylakoids (30 µg Chl/lane) isolated from wild type and TPK5-knock-out (left panel) and TPK3-knock-down (right panel) plants were loaded and assayed with the monoclonal antibody. The same membranes were stripped and reblotted with anti-ATP-ase to check for equal loading. E) Tonoplast and thylakoid fractions (20 µg of total protein of each) were loaded and developed with anti-TIP1.1 antibody (TIP1.1 is indicated by arrow at 28 kDa). In A, C and E nitrocellulose membranes and the BCIP/NBT (Sigma) development system, while in B and D PVDF membrane and ECL system was used.