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2655
  • Biotinylation of Antibody ( Contributed by Nanci Donacki )
      
  • Antibody labeling (House Ear Institute)
    Labeling protocols using affinity markers.
      
  • Coupling Colloidal Gold to Protein (Walter Steffen)

     

    • Gold labelling of rabbit IgG

       

    • Preparation of enzyme gold complex

       

    • Coupling of Nanogold to IgG
        

       

  • Production of Colloidal Gold (Walter Steffen)
      

     

  • Protein (Antibody) Fluorescent Labeling (Mario Roederer)
    Detailed protocol for conjugation of proteins (principally monoclonal antibodies) to fluorescent dyes. The conjugations fall into four basic protocols: Type 1 (used for FITC, Cy5, and the initial preparation of the Cy5 and Cy7 tandem dyes); Type 2 (used for Biotin and Cascade Blue); Type 3 (used for conjugation of PE, APC, TR-BSA, and their derivatives); and Type 4 (used for the initial preparation of non-immunoglobulin proteins like Annexin V). Procedures for the conjugation of immunoglobulins are provided for the following molecules:
    • Fluorescein (FITC)
    • Phycoerythrin (PE)
    • Cy5PE
    • Cy7PE
    • Texas Red (TR)
    • Allophycocyanin (APC)
    • Cy5
    • Cy7APC
    • Cascade Blue
    • Biotin

In addition, protocols for conjugation of Annexin V to the following molecules are given as models for fluorescent conjugation of non-immunoglobulin proteins. 

    • FITC
    • PE
    • TR-BSA
    • APC
    • Cascade Blue
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