Labeling Procedure for IgG1. Prepare 10 mM of the biotin labeling reagent using DMSO.2. Prepare 100 μl of 1 mg per ml IgG buffer solution (pH 7.5-8.5) that does not contain any large molecules with amine compounds.3. Add 1-5ml biotin labeling reagent DMSO solution to the IgG buffer solution and incubate at 37ºC for 1 hour.4. Remove excess biotin labeling reagent using gel filtration or dialysis.5. Prepare solutions for further experiment using an appropriate buffer such as PBST (0.05% Tween 20/PBS).
1. P. Kongtawelert and P. Ghosh, A New Sandwich-ELISA Method for the Determination of Keratan Sulphate Peptides in Biological Fluids Employing a Monoclonal Antibody and Labelled Avidin Biotin Technique., Clin. Chem. Acta,, 1990, 195, 17 .2. G. Paganelli, S. Pervez, A. G. Siccardi, G. Rowlinson, G. Deleide, F. Chiolerio, M. Malcovati, G. A. Scassllati and A. A. Epenetos, Intraperitoneal Radio-localization of Tumors Pre-targeted by Biotinylated Monoclonal Antibodies, Int. J. Cancer, 1990, 45, 1184.3. C. Wagener, U. Kruger and J. E. Shively, Selective Precipitation of Biotin-labeled Antigens or Monoclonal Antibodies by Avidin for Determining Epitope Specificities and Affinities in Solution-phase Assays, Methods Enzymol., 1990, 184, 518 .4. D. M. Boorsma, J. Van Bommel and E. M. Vander Raaij-Helmer, Simultaneous Immunoenzyme Double Labelling Using Two Different Enzymes Linked Directly to Monoclonal Antibodies or with Biotin-avidin, J. Microscopy, 1986, 143, 197.5. A. Komura, T. Tokuhisa, T. Nakagawa, A. Sasase, M. chihashi, S. Ferrone and Y.Mishima, Specific Killing of Human Melanoma Cells with an Efficient 10B-compound on Monoclonal Antibodies, Pigment Cell Res., 1989, 2, 259.6. R. Rappuoli, P. Leoncini, P. Tarli and P. Neri, Competitive Enzyme Immunoassay for Human Chorionic Somatomammotropin Using the Avidin-biotin System, Anal. Biochem., 1981, 118, 168.