Cy3-streptavidin

Fig. 3. Schematic of staining process of SARS-CoV immunochip. (1) Spotting A high-precision robot transfers the samples, SARS-CoV proteins, and glycans of various complexities, from 96-well plate to nitrocellulose-coated glass slides. (2) Staining Before staining, the slides are rinsed with IX phosphate-buffered saline (PBS), and blocked with 1% bovine serum albumin (BSA)-PBS containing 0.05% NaN3 and 0.05% Tween-20. They are subsequently incubated with horse anti-SARS sera. The primary antibodies captured by microarrays are detected using biotinated anti-horse immunoglobulin (Ig)G, and visualized by Cy3-streptavidin.

Biotinylated probe (B,G,N) AMCA-antibiotin (J) AMCA-streptavidin (J) AMCA-NeutraLite avidin (M) Cy2-antibiotm (J) Cy2-streptavidin (J,A) FITC-UltraAvidin (L) Cy3-antibiotin (J) Cy3-streptavidin (J,A) CyS-antibiotin (J) Cy5-streptavidin (J,A)... 

Tyramide signal amplification (TSA PerkinElmer Life Sciences, Boston) and enzyme-labeled fluorescence (ELF Molecular Probes) are related detection technologies. In the tyramide amplification process, a tyramide-biotin complex is produced by the action of horseradish peroxidase. The complex precipitates near the binding site and accumulates. The complex is detected by the use of streptavidin-Cy3/Cy5. 

Fig. 6. Measurement of the relative amount of ligand bound to each protein in the array. (A) Schematic of on-chip binding assay in which a fluorescently labeled interaction partner binds to the functional, arrayed protein immobilized to the streptavidin-coated surface via the biotinylated BCCP tag. (B) p53 protein function microarray probed with Cy3-labeled GADD45 duplex oligo. Quantification of the signal intensity from each spot allows the effect of polymorphic and functional variation on the DNA binding function of p53 to be determined.

Streptavidin-Cy3 and streptavidin-Cy5 conjugates (Amersham Pharmacia, Piscata-way, NJ). 

In a protein-binding assay with fluorescence detection a microarray of biotin, HPYPP-peptide and WSHHPQFEK-peptide was screened against streptavidin-Cy3 and avidin-Cy5. By following the same principle an anti-human insulin monoclonal antibody was also screened against a set of different peptides. 

To demonstrate that STORM can indeed resolve nearby fluorescent molecules with sub-diffraction-limit resolution, we first engineered samples with known relative positions of the fluorescent labels - double-stranded DNA labeled with two Cy3-Cy5 pairs separated by a well-defined number (135) of base pairs, corresponding to an inter-CyS distance of 46 nm along the contour of DNA [4]. The DNA strands were immobilized in a flat configuration to a quartz slide through multiple biotin-streptavidin linkages. The two Cy5 dyes were turned on and off, repetitively, and the image sequence was analyzed to determine the positions of individual activated Cy5 dye. We then constructed... 

Figure 25. Intuitive description of directional fluorescence emission from hybridized DNA. Figure not drawn to scale, BSA-streptavidin =90A, Cy3-DNA = 70A (adopted from [43]).

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