Conjugation via common conjugates at various scales
To be useful as detection and capture reagents, antibodies often are conjugated to fluorophores, enzymes, biotin and other such molecules. Similarly, recombinant proteins often are conjugated to allow them to efficiently couple with solid supports, or to improve their immunogenicity for antibody development.
- Used to label both antibodies and recombinant proteins for the purposes of coupling to streptavidin-coated substrates.
- Biotin-conjugates are useful for:
- immobilization of biotinylated antibody/protein to solid-phase supports (assay plates, beads, etc.); and
- labeling primary antibodies to allow recognition by Streptavidin conjugates for immunofluorescence, flow cytometry, Western blotting, etc.
- Used to label antibodies for use in fluorescence microscopy, cell sorting, etc.
- Used to label antibodies for the purpose of detection by Western blotting, ELISA, and Immunohistochemistry.
KLH (Key Limpet Hemocyanin)
- KLH is the protein most commonly used for conjugation to immunogens.
- Small molecular weight peptides, highly conserved and "self" proteins are generally non-immunogenic.
- Conjugation to KLH renders them immunogenic for the purpose of antibody production.
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