Attachment of a Biotinylated IgG/Elution of Purified Antigen 3. Preparation of Streptavidin-Coated Microspheres 2. Goat Anti-Mouse IgG-Coated Microspheres A.
These values allow for easy optimization of reagent requirements, without the added time and expense of a completely empirical determination of ligand concentration. Our streptavidin-coated microspheres are assayed in terms of their ability to bind biotinylated alkaline phosphatase, our protein A-coated microspheres are assayed for human IgG binding capacity, and our goat anti-mouse secondary antibodycoated microspheres are assayed for their capacity to bind mouse IgG. All of our protein-coated microspheres are formulated to optimize this orientation. For example, antibodies should be bound at their Fc (rather than Fab) region to maintain optimum biological activity. When binding biological ligands to solid supports such as microspheres, the orientation by which they bind is often crucial. Fc-directed attachment (and elution) of IgG to Protein A-coated microspheres (A)/ covalent cross-linking procedure (B) Preparation of ProActive® Protein A coated microspheres 2. Physical Parameters General Guidelines Sample Procedures 1. In addition, because the biotinylated ligand is set off from the surface of the microspheres, the steric hindrance that can cause a problem when coupling to functionalized microspheres is reduced, especially for larger ligands.
These will then bind directly to our streptavidin-coated microspheres with a bond strength approaching that of a covalent bond (Ka=1015/M). Virtually any biological ligand can be biotinylated through a onestep chemical reaction. These offer several advantages in terms of ligand attachment over traditional plain or surface-functionalized microspheres,īackground Streptavidin-and Biotin-Coated Microspheres A. Microspheres pre-conjugated to various types of generic binding proteins and secondary antibodies are rapidly becoming the solid phase support of choice in many areas, including immunological applications, nucleic acid work, and cell separation and visualization.
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