Enzyme-linked interaction assay competitive

Competitive Enzyme-Linked Interaction Assay (cELIA)... 

Consequently, in order to determine whether any of the combination of mutations did in fact include binding-site residues, the binding affinities of the mutated cohesins were also evaluated in a quantitative manner. The results are presented in Figure 1. In competitive enzyme-linked interaction assay, cELIA, the native cohesin was used as a standard to coat microtiter plates. The immobilized cohesin was then allowed to interact with an enzyme-linked dockerin solution together with a competitor cohesin (native or mutated) in the solution phase. The measured enzymatic activity, expressed as the percentage of activity detected in the absence of the soluble competitor, reflects die amount of dockerin bound to the immobilized cohesin standard. The IC50, i.e., the... 

Fig. 10.10. Determination of thermogenin amount in brown adipose tissue mitochondria by the enzyme-linked immunosorbent assay (ELISA) system. The amount of thermogenin was determined as elsewhere described (Cannon et al. [13] Sundin et al. [40] Hansen et al. [56]) in an assay system based on the competition between absorbed and added thermogenin for rabbit on/r-rat-thermogenin antibodies. The interaction was followed with a sheep onri-rabbit-IgG antibody conjugated to alkaline phosphatase. The reaction was linearized as indicated (abs 0 is the absorbance developed in the absence of competing thermogenin). It is seen that this assay can detect less than 0.25 fig thermogenin, i.e., the content in less than 10 fig of mitochondria. It is also seen that the thermogenin content of rat brown fat mitochondria is approximately doubled after a 24 h cold stress. (Our unpublished observations.)...

Antibodies can be used for a variety of applications in the molecular characterization of receptors and receptor-hgand interactions. Antibodies can be used for the detection of receptors in tissue shces. Western blot experiments [48], or ELISAs (enzyme-linked immunosorbent assays) [49]. They can also be used in competition experiments to map the binding epitope of a hgand [50]. Even though the use of antibodies is routine, fhere is no general protocol for fheir generation. 

Many types of assay are available to be used in HTS protocols to identify inhibitors of PPIs, but a competition assay, in which inhibition of complex formation is measured, is most common. Fluorescence polarization (FPA), fluorescence resonance energy transfer (FRET), enzyme-linked immunosorbent assays (ELISA), and other assay formats have been used. The interacting proteins can be used in their full-length forms though, more frequently only the interacting domains are employed, and if possible the excised interacting peptide is usually preferred. 

Immunoassay techniques are based on the antigen-antibody interaction. These techniques involve a competitive reaction between antigen molecules of the target molecules and labeled antigen molecules for a limited number of antibodies. Enzyme-linked immunosorbent assays (ELISA) in which antibodies are immobilized on a solid phase are the most popular for pesticide detection. As pesticides are small molecules, in order to synthesize antibodies, pesticide derivatives (haptens) must be synthesized and coupled to carrier proteins. ... 

The antagonistic effect of all 25 candidate compounds on LFA-l/ICAM-1 binding was tested in a competition enzyme-linked immunosorbent assay (ELISA). One weakly active compound was identified that blocked the interaction with an IC50 value of 70 [xM (compound 6). This compound was identified by similarity searching. 

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