The BabyBio ACT columns are prepacked with WorkBeads 40/1000 ACT pre-activated resin. This resin is a reliable starting material for the preparation of customized chromatography resins. A wide number of organic molecules and biomolecules containing sulfhydryl-, amino- or hydroxyl groups can be coupled covalently by nucleophilic displacement to the agarose matrix according to the well-known bromohydrin activation method (see Datasheet).
The main characteristics of BabyBio ACT columns are shown in the table below. For more detailed instructions of how to use BabyBio ACT, see the user instruction.
WorkBeads 40/1000 ACT is pre-activated to have a bromohydrin group that is reactive towards primary amines, thiol- (sulfhydryl), hydroxyl-, or histidyl residues. Most substances containing one or more of these groups can be coupled to the resin. The coupling reaction results in a stable covalent bond. The blocking step is required to eliminate further coupling of substances that are in contact with the resin during subsequent use of the prepared resin. Blocking is often carried out using ethanolamine or ß-mercaptoethanol by reaction of the amine or thiol group.
The bromohydrin coupling method does not introduce additional charges. The coupling is done at room temperature in an aqueous solution and does not release hazardous chemicals during normal use. The work can be carried out on the lab bench as long as the substance to be coupled is not hazardous.
Ligands with free amino and sulfhydryl groups will couple easily overnight in basic pH conditions at room temperature. Coupling of substances containing hydroxyl groups requires high pH (pH > 12) due to the low nucleophilicity of this functionality and the hydroxyl groups need to be deprotonated. Coupling of thiol-containing substances can be carried out under weakly alkaline conditions.