Guanosine- 5'- O- diphosphate, immobilized on a polymeric matrix (GDP-Gel / GDP-Agarose)

Suspension in 30 mM Na₂HPO₄ buffer (pH 7). Ligand density: approximately 37.2 umol/ml of settled gel UV:λ_max 252 nm/suspension in glycol.

GDP-Gel has sufficient stability for chromatography at ambient temperature and does not need special care during handling or shipment. Nevertheless, for longer storage periods the gel should be kept in the refrigerator at 4 - 8°C. Storage buffer should contain 0.1% sodium azide for prevention of microbial growth.

GDP-Gel is suitable for the affinity purification of fucosyltransferases.

After equilibration with 20 mM Mes-KOH, pH 6.8, containing 0.02% NaN3 and 1 mM dithioerythreitol the affinity column is loaded with the protein solution, e.g. at 1 mL/min. The column is then washed with 20 mM Mes-KOH buffer, pH 6.8, containing 50 mM NaCl, 0.02% NaN3, and 30% glycerol and subsequently with the same buffer containing 500 mM NaCl. Elution of the enzyme can be performed with the same buffer containing 1.5 M NaCl (all data according to Costa et al. 1997).
Regeneration of the gel may be achievable by incubation with 8 M urea or other chaotropic agents and subsequent washing with a suitable buffer.