Thermo Scientific Pierce Iminobiotin Agarose provides for unusual affinity purification experiments involving clean-up or removal of streptavidin or avidin protein components.
Pierce Iminobiotin Agarose is beaded agarose support onto which iminobiotin moieties have covalently immobilized. Typical avidin-biotin affinity procedures use immobilized avidin or streptavidin proteins to capture biotin-tagged molecules from a sample. Occasionally, however, the reverse situation arises in which immobilized biotin in needed to capture avidin or streptavidin components from a sample. This iminobiotin resin addresses this special situation.
- Enable purification or removal of avidin or streptavidin samples
- Crosslinked beaded agarose onto which iminobiotin is covalently immobilized
- Use Iminobiotin Agarose for reversible binding (e.g., non-denaturing purification of avidin)
Immobilized iminobiotin is the guanido analog of biotin. The dissociation constant of the avidin-iminobiotin complex is pH dependent. At pH 9.5 to 11, the avidin-iminobiotin complex will bind tightly. At pH 4, the avidin-iminobiotin complex will dissociate. Because no denaturing agents such as 8M guanidine hydrochloride or 4M urea are required for elution, the avidin conjugate has a better chance of maintaining its activity after purification. Thus, iminobiotin agarose can be used for situations requiring reversible binding and elution of avidin components.
|Chemical structure of Thermo Scientific Pierce Iminobiotin Agarose. Each porous agarose bead is 45 to 165µm diameter and contains trillions of iminobiotin groups.
References (for both biotin and iminobiotin):
- Chaiet, I. and Wolf, F.J. (1964). Arch. Biochem. Biophys. 106, 1-5.
- Green, N.M. (1975). Avidin. Advances in Protein Chemistry. New York: Academic Press, pp. 29, 85-133.
- Hofmann, K., Wood, S.W., Brinton, C.C., Montibeller, J.A. and Finn, F.M. (1980). Proc. Natl. Acad. Sci. 77(8), 4666-4668.
- Wood, G.S. and Warnke, R. (1981). J. Histochem. Cytochem. 29, 1196-1204.
- Gitlin, G., Bayer, E.A. and Wilchek, M. (1987). Biochem. J. 242, 923-926.
- Hiller, Y., Gershoni, J.M., Bayer, E.A. and Wilchek, M. (1987). Biochem. J. 248, 167-171.
- Alon, R., Bayer, E.A. and Wilchek, M. (1990). Biochem. Biophys. Res. Comm. 170, 1236-1241.
- Gao, C., et al. (1997) Proc. Natl. Acad. Sci. 94, 11777-11782.
- Pazy, Y., et al. (2003). J. Biol. Chem. 278(9), 7131-7134.
Review of affinity purification
Review of avidin-biotin interaction
Biotin Agarose (for irreversible binding)