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These mAb were analyzed for their fine specificity by measuring binding to synthetic analogs of the decapeptide, and cDNA sequences encoding the mAb V regions were determined.
Protocols are given for labeling with dyes and polyethylene glycol, for quantitatively measuring binding to cell surface receptors by kinetics and thermodynamics, and for exploiting new engineering opportunities from using "click chemistry" with nonnatural amino acids.
Adhesion to host receptors was determined by measuring binding to purified receptors coated onto Petri dishes.
Non-specific background adhesion was determined by measuring binding to Duffy antigen coated on Petri dishes.
First, the specificity and relative affinity to Aβ were tested by measuring binding to immobilised Aβ 1 40 peptide.
Serum IgG or IgM against dsDNA was detected by enzyme-linked immunosorbent assay (ELISA) by measuring binding to coated calf thymus DNA (Sigma).
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CETSA therefore provides a feasable alternative to measure binding to a target where traditional biophysical methods are not possible, for example when recombinant protein cannot be produced.
[3H]-triptolide was prepared and a filtration assay designed to measure binding to cells and cellular extracts.
Proteins in the reaction buffer were injected into the chamber to measure binding to DNA substrate.
First, we measured binding to the free 2F5 peptide captured on the ELISA plate (Fig 3B).
Second, and in parallel we measured binding to the ES4 protein target captured on the plate (Fig S1).
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