A new homogeneous competitive assay of ligand affinities was proposed based on quenching the fluorescence of tryptophan/tyrosine residues in a protein via Főrster-resonance-energy-transfer using a fluorescent reference ligand as the acceptor. Under excitation around 280 nm, the fluorescence of a protein or a bound acceptor was monitored upon competitive binding against a nonfluorescent candidat...

The 'sandwich' binding format, which uses two reagents that can bind simultaneously to a given analyte, is the gold standard in diagnostics and many biochemical techniques. One of the bottlenecks in creating a sandwich assay is identifying pairs of reagents that bind non-competitively to the target. To bridge this gap, we invented Megaprimer Shuffling for Tandem Affinity Reagents (MegaSTAR) to ...

A method is described for quantitation of testosterone in plasma from females and males. The principal operations of extraction, chromatographic fractionation, and competitive protein-binding assay can be completed for eight duplicate samples in a single day. Experimental data used in the development of the test and the rationale for the specific operations and conditions are presented. The spe...

A sensitive technique for plasma insulin assay based on displacement of labeled insulin from its antibody was reported by y ALLOW and BERSON in 1960 (9). Subsequently, similar methods based on competitive protein binding have been deve1oped. In 1963 MURPHY and co-workers (6) described a technique involving a dialysis using 14C-cortisol. The time required for this procedure was shortened from tw...

The predictions made for fold recognition and modeling accuracy at the 1996 Critical Assessment of Structure Prediction meeting (CASP2) were assessed to discover which groups did best. With 32 groups making a total of 369 predictions, it was necessary to use simple criteria for distinguishing between the entries. By focusing on the predictors' ability to use the sequence of the unknown target s...

BACKGROUND Streptococcal protein G comprises two or three domains that bind to the constant Fc region of most mammalian immunoglobulin Gs (IgGs). Protein G is functionally related to staphylococcal protein A, with which it shares neither sequence nor structural homology. RESULTS To understand the competitive binding of these two proteins to the Fc region, the crystal structure of a single Ig-...

Competitive adsorption of three human plasma proteins: albumin (HSA), fibrinogen (Fgn), and immunoglobulin G (IgG) from their ternary solution mixtures onto a sulfhydryl-to-sulfonate gradient surface was investigated using spatially-resolved total internal reflection fluorescence (TIRF) and autoradiography. The concentration of each protein in the ternary solution mixture was kept at an equival...

Targeting sites that modulate protein-protein interactions represents an ongoing challenge for drug discovery. We have devised an assay principle, named ligand-regulated competition (LiReC), in an effort to find non-ATP competitive small-molecule regulators for type Ialpha cAMP-dependent Protein kinase (PKA-Ialpha), a protein complex that is implicated in disease. Our assay based on the LiReC p...

Solid-phase binding, competitive binding, and cytotoxicity neutralization assays indicate that the B pentamer and A subunit both contribute to human serum amyloid P (HuSAP) component binding to Stx2. A polyvalent globotriaosyl-ceramide receptor analog, Daisy, did not competitively inhibit HuSAP binding, implying that the two ligands bind to different Stx2 domains.