Recombinant biotin-binding phages were affinity-selected from a random peptide library expressed on the surface of filamentous phage. Phage binding to biotinylated proteins was half-maximally inhibited by micromolar concentrations of a monobiotinylated molecule. Sequencing of the peptide inserts of selected phages led to the identification of a previously unknown biotin-binding motif, CXWXPPF(K...

The Serial Analysis of Gene Expression (SAGE) method, described in 1995 by Velculescu et al ., represents a powerful means to compare gene expression between two mRNA populations. An improvement to SAGE that removes contaminating linker molecules, which compromise the efficiency of the method, has been developed. This modification utilises biotinylated PCR primers, which generate biotinylated l...

Proximity ligation assay (PLA) is a recently developed strategy for protein analysis in which antibody-based detection of a target protein via a DNA ligation reaction of oligonucleotides linked to the antibodies results in the formation of an amplifiable DNA strand suitable for analysis. Here we describe a faster and more cost-effective strategy to construct the antibody-based proximity ligatio...

The Ag nanoparticle based localized surface plasmon resonance (LSPR) nanosensor yields ultrasensitive biodetection with extremely simple, small, light, robust, and low-cost instrumentation. Using LSPR spectroscopy, the model system, biotinylated surface-confined Ag nanotriangles, was used to detect less than one picomolar up to micromolar concentrations of streptavidin. Additionally, the monito...

Vol. 40, No. 5 (2006) The binding of avidin to biotin is the strongest known noncovalent biological interaction [dissociation constants (Kd) approximately 10 -15 M] between a protein and ligand (1). The high affinity and specificity of the avidin-biotin interaction is utilized for enriching biotin-containing proteins from mixtures (2,3). Water-soluble Nhydroxysulfosuccinimide (sulfo-NHS) deriva...

Monodisperse biotinylated poly(styrene-co-N-acryloxysuccinimide) microspheres were synthesized in aqueous solutions with a two-step method. Upon the addition of avidin solutions of different concentrations in phosphate buffer into the dispersed biotinylated microspheres, the microspheres aggregated rapidly due to the high binding affinity between biotin and avidin. The hydrodynamic diameter of ...

The attachment of biotin to a small molecule provides a powerful tool in biology. Here, we present a systematic approach to identify biotinylated analogues of the Hsp90 inhibitor PU-H71 that are capable of permeating cell membranes so as to enable the investigation of Hsp90 complexes in live cells. The identified derivative 2g can isolate Hsp90 through affinity purification and, as we show, rep...

3-Methylcrotonyl-CoA carboxylase (MCCase) is a nuclear-encoded, mitochondriallocalized biotin-containing enzyme. The reaction catalyzed by this enzyme is required for leucine (Leu) catabolism in mitochondria, and it may also play a role in the catabolism of isoprenoids and in the mevalonate shunt. In Arabidopsis, two MCCase subunits (the biotinylated MCCA subunit and the non-biotinylated MCCB s...

biotinylated antibody bound to streptavidin-coated donor beads and a second antibody conjugated to AlphaLISA acceptor beads. The binding of the two antibodies to the analyte brings donor and acceptor beads into proximity. Laser irradiation of donor beads at 680 nm generates a flow of singlet oxygen, triggering a cascade of chemical events in nearby acceptor beads, which results in a chemilumine...

Cyclic polyolefin (COP) is an inexpensive hydrophobic material with low auto-fluorescence, high light transmittance and thermal stability, broad chemical resistance and no non-specific protein binding. Here, the hydrophobic alkane COP was modified to have carbonyl functionalities through oxygen plasma and chemical etching treatments to increase usefulness for chemical and biochemical applicatio...