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Cibacron Blue - Agarose

Catalog Number: 191263
Cibacron Blue - Agarose


Structure:



Molecular Formula: C29H20N7O11S3Cl

Molecular Weight: 774.168

Ligand: Contains 2-5 umoles of Cibacron Blue F3GA per ml gel. 4% crosslinked agarose; molecular exclusion limit is approximately 20 million dalton. Specific ligand concentration is determined by aliquot of resin suspended in 75% glycerol andn OD260 measured against unsubstituted agarose beads.

Bead Size Distribution: Approximately 60 - 160 um.

Form: Suspension in distilled water containing 20% ethanol as a preservative.

Applications: Useful in the isolation of enzymes requiring NAD+ and NADP, albumin, interferon, steroid receptors, and 1,25-Dihydroxyvitamin D3-receptor.

Description: Certain reactive textile dyes have been found to bind to proteins especially those with affinities to various nucleotides.3,8 Immobilized dyes have been found to bind from 5 to 60% of the proteins in various crude cell extracts.11 The affinity of a specific dye for a particular nucleotide binding site on a protein cannot be predicted. The affinity for reactive dyes to proteins may be due to substrate/cofactor similarities as well as hydrophobic and ion exchange properties. Some proteins have been found to require the addition of divalent cations for binding to dye resins.5 Nonionic detergents have been found to encapsulate immobilized dyes in micelles and prevent proteins from binding. Low concentrations of anionic detergents will be repelled by the negative charge on most reactive dyes and may not interfere with protein binding.10

Cibacron blue has been shown to bind to several enzymes with known affinities to nucleotide cofactors. It has also been shown to bind to dehydrogenases9, kinases7,13, restriction endonucleases1, albumin14, and interferon.6

Typical Procedure:

Suggested Conditions for Use:

Protein determination should be performed on the sample to be loaded as well as the wash and eluant fractions. Protein binding capacity will vary greatly and can exceed 20 mg per ml of resin. Most steps can be done either in a column or batchwise using a filter funnel or centrifugation. Care should be taken to prevent the media from drying out completely.


Regeneration and Storage: Wash suspension with approximately 5-10 column volumes of each solution.

References:
  1. Baksi, K., et al., Biochemistry, v. 17, 4136 (1978).
  2. Burton, S.J., et al., J. Chromatog., v. 435, 127 (1988).
  3. Haeckel, R., et al., Hoppe-Seyler's Z. Physiol. Chem., v. 349, 699 (1968).
  4. Hanggi, D. and Carr, P., Anal. Biochem., v. 149, 91 (1985).
  5. Hughes, P., et al., Biochim. Biophys. Acta, v. 700, 90 (1982).
  6. Jankowski, W.J., et al., Biochemistry, v. 15, 5182 (1976).
  7. Kobayashi, R. and Fang, V.S., Biochem. Biophys. Res. Commun., v. 69, 1080 (1976).
  8. Kopperschlager, G., et al., FEBS Lett., v. 1, 137 (1968).
  9. Lamkin, G.E. and King, E.E., Biochem. Biophys. Res. Commun., v. 72, 560 (1976).
  10. Robinson, J.B., et al., Proc. Natl. Acad. Sci. USA, v. 77, 5847 (1980).
  11. Scopes, R.K., J. Chromatog., v. 376, 131 (1986).
  12. Stellwagen, E., Meth. Enzymol., v. 182, 343 (1990).
  13. Thompson, S.T., et al., Proc. Nat. Acad. Sci. USA, v. 72, 669 (1975).
  14. Travis, J., et al., Biochem. J., v. 157, 301 (1976).