Bio-Nobile immobilized metal affinity chromatography (IMAC)

QuickPick IMAC magnetic particles are formed from magnetizable agarose particles which affords an extensive porous surface area especially preferred in the purification of histidine tagged proteins from cell lysates. The transfer of magnetic particles with the QuicPick magnetic tool enables rapid purification of histidine tagged proteins with suitable binding and elution characteristics. The methodology used abolishes the need for sample pre-treatment or use of time-consuming column technologies.

High quality purifications using QuickPick™ IMAC

The performance of the QuickPick IMAC kit in purifying His-tagged protein specifically is illustrated by the example below in which E.coli His-tagged glutathione-S-transferase (HT-GST) was isolated using the QuickPick™ IMAC kit. The cells were lysed in 20 mM Tris-HCl buffer, pH 7.0 before purification.
Download sample QuickPick™ IMAC purification protocol
M 1 2 3 4 5
kDa
97
66
45
30
20
14
M = Molecular weight marker
1 = Cell lysate
2 = Lysate after MP treatment
3 = 5 µl QuickPick IMAC eluate
4 = 7.5 µl QuickPick IMAC eluate
5 = 10 µl QuickPick IMAC eluate

Purity checked by analyzing 5, 7.5 and 10 µl of eluate on SDS-PAGE gel. The HT-GST protein band is clearly seen for each of the three eluate lines.

Higher yields by using the QuickPick™ IMAC Plus kit

The QuicPick IMAC Plus kit is designed for applications which demand higher yields of protein. As shown below, a yield of about 0.5 mg (~30 kDa protein) is obtainable from a single purification.
Yields obtained using QuickPick™ IMAC and IMAC Plus kits
IMAC IMAC Plus
Sample material
Bacterial cells E. coli recombinant protein expression:
Cells from 5 ml cultivation lysed in 500 µl wash buffer		 E. coli recombinant protein expression:
Cells from 10-40 ml cultivation lysed in 3 ml wash buffer
Sample volume 300-1000 µl 2-4 ml
Processing time for
- one sample 10 min 10 min
- four samples 30-40 min 30-40 min
Elution volume per samle 20-100 µl 300-600 µl
Yield 30 ± 3 µg for HT-GST Up to 0.5 mg for HT-GST