Streptavidin phycoerythrin

The spectral properties of four major phycobiliproteins used as fluorescent labels can be found in Tables 9.1 and 9.2. The bilin content of these proteins ranges from a low of four prosthetic groups in C-phycocyanin to the 34 groups of B- and R-phycoerythrin. Phycoerythrin derivatives, therefore, can be used to create the most intensely fluorescent probes possible using these proteins. The fluorescent yield of the most luminescent phycobiliprotein molecule is equivalent to about 30 fluoresceins or 100 rhodamine molecules. Streptavidin-phycoerythrin conjugates, for example, have been used to detect as little as 100 biotinylated antibodies bound to receptor proteins per cell (Zola et al., 1990). 

The effects of CVS on the numbers of T-cell subsets were examined on day 9 at the time of tumor i.v. rechallenging, Fig. (6)-Protocol B. Cells were stained with fluorescein 5-isothiocyanate (FITC)-, phycoerythrin (PE)-conjugated and/or biotinylated mAb and were additionally stained with RED-613-streptavidin for flow-cytometric three-color analysis. 

Fig. 5.2. Methods for detection of passenger-ligand interaction. (A) Fluorophore-coupled ligand (B) fluorophore-coupled antibody (C) quaternary complex generated by subsequent rounds of incubation with ligand, primary antibody, biotinylated second antibody, and strep ta-vidin, R-phycoerythrin conjugate (D) quaternary complex generated by subsequent rounds of incubation wi tli ligand, primary antibody, biotinylated second antibody, and streptavidin-coated magnetobeads. L ligand P passenger.

Streptavidin, R-phycoerythrin conjugate, lmgmL-1 (Molecular Probes). 

If the ligand protein is fluorescently labeled, resuspend the cells in 10 pL PBS and subject directly to FACS. If the ligand protein is biotinylated (protocol 2), resuspend the cells in 10 pL PBS containing streptavidin, R-phycoerythrin conjugate (1 10 dilution in PBS). Incubate on ice again for 10 min. 

Naegleria fowleri (Pougnard et al. 2002) has been detected by SPC in surface water. A monoclonal antibody was conjugated with biotin or HRP and revealed by streptavidin conjugated to RPE-Cy5 (R-phycoerythrin conjugated with the cyanine dye Cy5) or FITC-conjngated tyramide, respectively. The RPE-Cy5 protocol was the most efficient and allowed the detection of both trophozoite and cyst forms in water. 

R-Phycoerythrin Streptavidin (Molecular Probes/Invitrogen Life Technologies). 

Samuelson, L.A., D.L. Kaplan, J.O. Lim, M. Kamath, K.A. Marx, and S.K. Tripathy. 1994. Molecular recognition between a biotinylated polythiophene copolymer and phycoerythrin utilizing the biotin-streptavidin interaction. Thin Solid Films 242 50-55. 

Biotinylated copolymers of 3-undecylthiophene and 3-thienylmethanoI using streptavidin as a cross-linker protein is an electroactive matrix for the attachment of a photoactive protein, phycoerythrin. The biotinylation of the copolymer improves the film forming properties and results in a stable mono-layer. The phycoerythrin binding to the biotinylated copolymer monolayer can be monitored through fluorescence microscopy at the air-water interface [451]. 

Figu re 4.8 Flow cytometry analysis of silica nanoparticles surface-modified with proteins. MPMS nanoparticles (a, c) and TEOS nanoparticles (b, d) modified with GFP (a, b) or phycoerythrin-conjugated streptavidin... 

Phycoerythrin Optotransduction Avidm/streptavidin-biotinylated lipids Optical 70... 

Avidin-biotin systems. Stable and highly active protein-LB/LS film composites can be accessed using avidin-biotin conjugation techniques. Thus, sensor elements displaying biotinylated GOx and phycoerythrin, and streptavidin and av-idin conjugates of phycoerj tiirin, have been bound to avidin-lipid and biotinylated lipid-streptavidin LB films, or biotinylated lipid layers have been produced [71-73]. 

---