ATP (Agarose) 4B-CL (Lyophilized)
Adenosine 5`-triphosphate (ATP)-Separopore® 4B-CL is used in the purification of ATP binding proteins. Any protein with an accessible ATP-binding domain will bind to ATP- Separopore® 4B-CL, regardless of activation state. The activated proteins will bind to ATP-Separopore® 4B-CL with higher affinity than the corresponding inactivated protein. This is important when purifying proteins from crude fractions where protein concentration is low, as this prevents non-specific binding. Better recovery and yields may be obtained when the ATP-Separopore® 4B-CL purification is followed by ion exchange chromatography.
The production method involves alkylation of the nucleotide, ATP followed by alkaline rearrangement to yield the corresponding N6-carboxymethyl derivative with subsequent condensation using 1,6-diaminohexane to give N6-[(6-aminohexyl)carbamoylmethyl)].
- Ligand: ATP
- Matrix: Separapore® 4B-CL (crosslinked agarose beads, 4 %)
- Particle size range: 52 – 165 μm
- Matrix activation: Cyanogen bromide
- Ligand density: 2 – 5 ATP μmol / ml drained gel
- Attachment: N6 of the purine ring
- Spacer: 11 atoms
- Supplied as lyophilized powder, 1g powder yields 8 – 10 ml of gel