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Although Myr p 2a comprises approximately half of the protein content of JJA venom [6], it is highly basic and may have a lower affinity for the solid phase than other venom components, which may result in a lower proportion of it binding to the solid phase compared to what is present in whole venom.
We have used alternating surfaces of magnetic beads and plastic immunotubes to avoid selection of phage binding to the solid support.
On one hand, the dissociated carboxyl groups of the polymer, which undergo the direct binding to the solid surface cause increase of the solid surface charge (usually anionic compounds adsorption forces the formation of positively charged groups on the surface).
We minimized non-specific binding by using fragments (Fab, Fab', or Fab'2) as capture antibodies and by using proprietary buffer reagents (in addition to the standard casein, BSA, and detergents) to minimize non-specific binding to the solid phase.
After incubation and washing away unbound gliadin and peptides, a secondary antibody binding to the solid phase antibody-antigen complex was added, and the amount of complex quantified by means of the tag on the secondary antibody.
However, the fact that similarly purified PR3 is reactive with such sera if binding to the solid phase is mediated by a particular anti-PR3 mouse monoclonal antibody makes it more likely that at least one epitope on the PR3 molecule is hidden upon adsorption to the plastic surface [ 28].
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Bisphosphonates inhibit calcification by binding to the surface of solid calcium phosphate crystals and acting as crystal poisons affecting both crystal growth and dissolution [ 47].
The absorption efficiency of the gold particles and bubble formation threshold might be changed as well due to the binding to the silica particles which provide a solid interface.
To separate phages binding to the ligand from non-binders, the ligand is immobilized on a solid carrier allowing for easy separation of the liquid phase, which contains predominantly non-binding phages, from the solid phase, which contains mainly binding phages.
Competitive ELISA showed that the antibody-binding capacity to the solid phase was reduced with 25%63%%, dependent on the amount of antigen added (1.0 5.0 ng/ml) to the diluted serum before application on the microtiter plate.
Competitive ELISA regarding LH showed that the antibody-binding capacity to the solid phase was reduced with 43%–87%, dependent on the amount of antigen added (1.0 5.0 ng/ml) to the diluted serum before application on the microtiter plate.
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