Figure 4

Application of the scFvs in ELISA and WB. A) Selected scFvs were tested by ELISA against the cell free-expressed proteins, E. coli-derived GFP and scFv-Trx 3xFlag proteins, using BSA and GST as negative controls. B) Intensity of the fluorescence signal of the GFP or Trx-specific scFvs in comparison to a scFv control signal or buffer obtained from the normalized data of the microarrays after incubation with 647Alexa Fluor labelled-GFP, 647Alexa Fluor labelled-Trx or anti c-myc followed by 555 AlexaFluor anti-mouse IgG, respectively. C) Different amounts of GFP or scFv-Trx were separated by SDS-PAGE and transferred to nitrocellulose membranes to determine the sensitivity of the anti-GFP and the anti-Trx scFvs (1:10 diluted) as primary antibodies. They were followed by an anti-c-myc tag and peroxidase-labeled anti-mouse IgG antibody, respectively. An anti-His6 mAb or an anti-M2 Flag mAb were used as positive controls to detect the target proteins.