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Fig. 3 | Biological Procedures Online

Fig. 3

From: Production of Monoclonal Antibodies against Human Trefoil Factor 3 and Development of a Modified-Sandwich ELISA for Detection of Trefoil Factor 3 Homodimer in Saliva

Fig. 3

Schematic diagram demonstrating the principle behind the developed sandwich ELISA detecting dimeric TFF3. a The steps of the developed sandwich ELISA: (a). First, anti-TFF3 mAb is coated on the ELISA plate. b. Human recombinant TFF3 or saliva is added. The TFF3 present in the sample binds to the mAbs. c. The FITC-conjugated secondary anti-TFF3 mAb (which is the same clone of the coated mAb) is added. d. HRP-conjugated anti-FITC antibody is added and there is a reaction to the FITC-labeled anti-TFF3 mAb. e. Lastly, the TMB substrate is added and the color develops. The intensity of the color is proportional to the amount of the dimeric TFF3 present in the sample. Dimeric TFF3 can bind to both the coated mAb and the secondary mAb and the colorimetric signal can be developed by the reaction of the HRP-conjugated anti-FITC antibody with the TMB substrate. b In contrast, the secondary mAb cannot bind to monomeric TFF3 which cannot develop the colorimetric signal

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