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. 2007 Feb;143(2):670–683. doi: 10.1104/pp.106.090159

Figure 4.

Figure 4.

Quality control of recombinant 14-3-3 protein and R18 competing peptide. A, Recombinant His-tagged protein was prepared of all five barley 14-3-3 isoforms. Two micrograms of each isoform was run on SDS-PAGE and stained with Bio-Rad BioSafe colloidal Coomassie. B, Functionality of the recombinant Hv14-3-3C protein was tested using the inhibition of 14-3-3 proteins on NR. NR was semipurified from barley leaf extract. The affinity of Hv14-3-3C for NR was tested using different concentrations of recombinant Hv14-3-3C, ranging from 1 to 25 nm (molecular mass 14-3-3C = 29.9 kD). C, Functionality of the competing peptide R18 was tested using 10 nm of Hv14-3-3C inhibition on the NR enzyme. Concentration range of the R18 peptide was between 10 nm and 20 μm.