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 Pierce Avidin Agarose can be used in a variety of applications for the affinity purification of biotinylated macromolecules. In one variation, an antibody that has an affinity for a particular antigen is labeled with biotin. Cells containing the antigen are lysed, then incubated with the biotinylated antibody to form a typical antigen/antibody complex. To isolate the antigen, the crude mixture is passed through an immobilized avidin or streptavidin column, which will bind the complex. After appropriate washes, the antigen can then be eluted from the column with a pH 2.8 elution buffer. The biotinylated antibody is retained by the column.
Highlights
- Strong , nearly irreversible biotin:avidin interaction
- Binding biotinylated anti-transferrinfrom serum (1)
- Binding biotinyl peptides and elution with a SDS/Urea solution (2)
- Hybridization of biotinylated RNA to its complementary DNA and binding to immobilized avidin, with subsequent elution of the single-stranded DNA (3)
- Purification of double-stranded DNA (4)
- Approximate Binding Capacity >20 µg biotin/ml gel
References
- Manning, J., Pellegrini, M. and Davidson, N. (1977). Biochemistry 16, 1364-1370.
- Pellegrini, M., Holmes, D.S. and Manning, J. (1977). Nucl. Acids Res. 4, 2961-2973.
- Swack, J.A., Zander, G.L. and Utter, M.F. (1978). Anal. Biochem. 87, 114-126.
- Wilchek, M. and Bayer, E.A. (1989). Protein Recognition of Immobilized Ligands. Hutchins, T.W., ed. Alan R. Liss, Inc., pp. 83-90.
- Sharma, K.K., et al. (2000) J. Biol. Chem. 275, 3767-3771.
- Claypool, S.M., et al. (2002) J. Biol. Chem. 277, 28038-28050.
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