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A general prerequisite for a fusion protein/target membrane binding assay is a soluble and correctly oligomeric form of the viral fusion protein ectodomain.
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The ER binding assay was a good predictor for the other two assay results when the antiestrogens were excluded (r(2) is 0.78 for the yeast assays and 0.85 for the E-SCREEN assays).
The ligand binding assay was a variant of the ELISA in which dilutions of ligand (rhFGF-2) were allowed to bind to a ligand binding protein such as rhPln.D1 that had been coated onto the wells in PBS-N3.
The reasons for this difference may be the presence of significantly large number of protein "targets" in the nuclear extract preparations utilized in the DNA-binding assay and the potential that disrupting preformed Stat3:Stat3 dimers, as in the DNA-binding assay is a more challenging task, as has previously been suggested.
Ligand binding assay is associated with a multiplicity of specificity issues (Findlay, 2008).
In rosette assay, samples are spotted on a membrane, and the binding assay is carried out in a 96-well plate [ 7].
The virus binding assay was performed using a previously reported protocol, with modifications (Qing et al., 2016).
The radioactivity of the binding assay was counted with a WIZARD2 automatic γ-counter from Perkin Elmer (Boston, MA).
Since the pharmacological properties (experimental radioligand binding assay) are variables with a wide numerical distribution, for plotting purposes, we have reported them as −log of the K i or IC50 (i.e. as pK i or pIC50).
The binding assay was performed with a constant flow rate of 20 µl/min at 25°C through the chip.
Results of the complement binding assay are presented as a mean fluorescence index [FI (Fluorescent Index), proportion of positive bacteria expressed as a percentage multiplied by the geometric mean MFI] in arbitary units [12], [17], [18].
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