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Identification of hepatic protein-protein interaction targets for betaine homocysteine S-methyltransferase

Fig 2

Elution profiles from control, intein and intein-BHMT loaded chitin columns.

The figure shows representative A280 elution profiles from control (beads only; 4 ml), intein (4 ml) and intein-BHMT (1 ml) loaded chitin columns, as well as representative Coomassie Blue stained SDS-PAGE gels of liver cytosol, flowthroughs loaded on these columns and samples of the eluted peaks. (A) Control columns were loaded with liver cytosol and eluted with a NaCl gradient. (B) The flowthrough from the control column was loaded onto the intein column and elution performed with the same salt gradient. (C) The flowthrough of the intein column was loaded onto the intein-BHMT column and elution carried out with a NaCl gradient. (D) SDS-PAGE of the protein fractions loaded onto the columns: liver cytosol (2 μl); flowthrough of the chitin column (2 μl); flowthrough of the intein column (2 μl); and flowthrough of the intein BHMT column (2 μl). (E) Eluted proteins using salt gradients and 2-mercaptoethanol (2-ME) excision were collected and analyzed by SDS-PAGE: chitin peak (40 μl); intein peak (40 μl); and intein-BHMT peak (40 μl). Molecular weight standards are shown on the right side of each gel.

Fig 2

doi: https://doi.org/10.1371/journal.pone.0199472.g002