Immunoblot analysis
To check if the transgenic plants had the expected changes in their
protein expression, we did western blot analysis. Total protein was
isolated from 4-week-old FbβCA3x lines. Equal loading of the
protein (25 µg) was checked by running 15% SDS-PAGE (Fig. 2d).Polyclonal antibodies, raised against FbβCA3 protein, were used
to immuno-detect the CA protein in different FbβCA3x lines(Fig. 2e). In the transgenic lines, the CA protein
(~28 kD) expression was 1.3 to 2.1 fold higher than in
the VC (Fig. 2f) . The CA protein was also present in the VC and
showed the expected band in the blot, which was due to the high homology
of AtβCA’s with FbβCA3 .