The instrument detects this fluorescent signal and creates a digital image of your sample. If there is sufficient target, the fluoresceinated secondary antibody alone yields a strong signal. If not, amplify the signal by incubating the blot with the anti-fluorescein tertiary antibody conjugated to alkaline phosphatase, followed by the substrate. The substrate is cleaved by alkaline phosphatase to yield additional fluorescent signal. Figure 2a Protein detection with Vistra ECF .
Alkaline phosphatase is conjugated to an antifluorescein tertiary antibody.
Protein detection with an alkaline phosphatase-conjugated secondary antibody. Nucleic acid detection with Vistra ECF . A hapten, such as fluorescein, is incorporated into the DNA probe. The hapten is then bound by an antibody conjugated to alkaline phosphatase. The chemifluorescence advantage Specificity and versatility.