SpringerProtocols: Abstract: 11. Conjugation of Antibodies to Biotin

11. Conjugation of Antibodies to Biotin

By: James D. Hirsch², Rosaria P. Haugland²

Abstract

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Antibodies can be conjugated to biotin by a number of chemical means. They can then be used in immunochemical procedures in conjunction with secondary reagents coupled to biotin-binding protein proteins such as avidin.

Affiliation(s): (2) Molecular Probes Inc, Eugene, OR

Book Title: Immunochemical Protocols: Third Edition

Series: Methods in Molecular Biology | Volume: 295 | Pub. Date: Dec-13-2004 | Page Range: 135-154 | DOI: 10.1385/1-59259-873-0:135

Subject: Immunology

Key Words: Biotin - Avidin - Antibody - Labeling - Affinity - Fab Fragments - Fluorescence

References

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1.	Haugland, R. P. (2002) Handbook of Fluorescent Probes and Research Products, 9th ed. Molecular Probes, Inc., Eugene, OR, pp. 119–134.

3.	Hermanson, G. T. (1996) Avidin-Biotin Systems, in Bioconjugate Techniques, Academic Press, San Diego, CA, pp. 570–592.

4.	Aslam, M. and Dent, A. (1998) Bioconjugation. Protein Coupling Techniques for the Biomedical Sciences. Grove%s Dictionaries Inc., New York, NY.

5.	Hirsch, J. D., Eslamizar, L., Filanoski, B. J., Malekzadeh, N., Haugland, R. P., Beechem, J. M., and Haugland, R. P. (2002) Easily reversible desthiobiotin binding to streptavidin, avidin, and other biotin-binding proteins: uses for protein labeling, detection and isolation. Anal. Biochem. 308, 343–357.

6.	Haugland, R. P. and You, W. W. (1996) Coupling of antibodies with biotin, in The Protein Protocols Handbook. (Walker, J. M., ed.), Humana Press, Totowa, NJ, pp. 293–301.

7.	Haugland, R. (2000) Antibody conjugates for cell biology, in Current Protocols in Cell Biology, vol. 2 (Bonifacino, J., Dasso, M., Harford, J., Lippincott-Schwartz, J., and Yamada, K., eds.) John Wiley & Sons, New York, NY, pp. 16.5.1–16.5.22.

8.	Fudem-Goldin, B. and Orr, G. (1990) 2-iminobiotin-containing reagent and affinity columns, in Methods in Enzymology, vol. 184 (Wilchek, M. and Bayer, E., eds.), Academic Press, New York, NY, pp. 167–173.

9.	Der-Balian, G. P., Gomez, B., Masino, R. S., and Parce, J. W. (1990) A fluorometric method for determining the degree of biotinylation of proteins. J. Immunol. Methods. 126, 281–285.

10.	Rao, S. V., Anderson, K. W., and Bachas, L. G. (1997) Determination of the extent of protein biotinylations by fluorescence binding assay. Bioconjugate Chem. 8, 94–98.

11.	Livianiou, E., Costopoulou, D., Vassiliadou, I., Leondios, L., Nyalala, J. O., Ithakissios, D. S., et al. (2000) Analytical techniques for determining biotin. J. Chromatog. A. 881, 331–343.

12.	Green, N. (1965) A spectrophotometric assay for avidin and biotin based on binding of dyes by avidin. Biochem. J. 94, 23c–24c.

13.	Page, M. and Thorpe, R. (1996) Purification of IgG using protein A and protein G, in The Protein Protocols Handbook. (Walker, J. M., ed.) Humana Press, Totowa, NJ, pp. 733–734.

14.	Page, M. and Thorpe. R. (1996) Purification of IgG using affinity chromatography on antigen-ligand columns, in The Protein Protocols Handbook. (Walker, J. M., ed.) Humana Press, Totowa, NJ, pp. 739–741.

15.	Hermanson, G. T., Mallia, A. K., and Smith, P. K. (1992) Immobilized Affinity Ligand Techniques. Academic Press, San Diego, CA.

16.	Andrew, S. M. and Titus, J. A. (2000) Fragmentation of immunoglobulin G, in Current Protocols in Cell Biology, vol. 2) (Bonifacino, J., Dasso, M., Harford, J., Lippincott-Schwartz, J., and Yamada, K., eds.) John Wiley & Sons, New York, NY, pp. 16.4.1–16.4.10.

17.	Lacey, E. and Grant, W. N. (1987) Photobiotin as a sensitive probe for protein labeling. Anal. Biochem. 163, 151–158.

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