Figure 6From: Antibody degradation in tobacco plants: a predominantly apoplastic processPulse chase analysis of antibody fragments in MAb 2G12 transgenic plant cells. Protoplasts from leaves of transgenic tobacco plants expressing the MAb 2G12 were washed and analysed by SDS-PAGE and western blotting (Panels C and D) or pulse-labelled for 1 hr and chased for the indicated periods of time (Panels A and B). Extracellular medium samples and homogenised cells were subjected to immunoprecipitation with Protein G/Protein A mix. Immunoprecipitated proteins were analyzed by SDS-PAGE and radiography. The asterisk corresponds to the fully assembled 2G12 antibody, while the lower case letters (a to i) indicate antibody fragments. Panels A and C were performed under non-reducing conditions, whereas samples in Panels B and D were reduced with the addition of 5% β-mercaptoethanol prior to boiling.Back to article page