His-Tag Purification
His-Tag Purification
Immobilized Metal Affinity Chromatography
(IMAC) is based on the interaction between certain superficial protein
residues (histidines, cysteines and to a lesser extent tryptophans) with
transition metal cations, forming chelated complexes.
These transition
metals are bound through a chemical reaction to the agarose beads, giving the
agarose an activation degree.
SERVA Nickel NTA Agarose Resin consists
of crosslinked agarose derivatized with nitrilotriacetic acid and loaded with
divalent nickel ions. The four metal-binding sites on the chelate enable high
protein binding and result in minimal metal leaching. This makes the resin
ideal for purification under reducing conditions.
Ni-Extrachel Agarose Resin has a polychelator ligand covalently coupled to a highly crosslinked agarose resin and is loaded with nickel ions. The resin works in presence of EDTA, DTT and other chemicals, which result in stripping of the metal ions with standard Ni-NTA or -IDA resins. Its specificity and stability allows a one-step purification without the need of pretreatment of samples for removal of nickel stripping agents.
SERVA Ni-NTA Magnetic Beads allow the rapid and easy small scale purification of histidine tagged proteins.
SERVA
IDA Agarose Resins have iminodiacetic acid groups covalently coupled to
crosslinked agarose beads.
This resin is loaded with a divalent metal (Ni2+,
Co2+). In comparison with other chelating resins such as
NTA-agarose, IDA has three sites available for interaction with metal ions,
instead of four with NTA. Therefore IDA resins are usually more easily
regenerated allowing an easier elution of bound proteins with lower
concentrations of imidazole.
The choice of resin depends on whether binding
capacity or selectivity is more important for purification and whether the
protein is easy or difficult to separate.
For easy column purifications with our bulk resins we offer empty columns for small volumes of 50 – 100 µl and 200 – 250 µl or larger volumes of 0.5 – 2 ml and 2 – 6 ml.
The
innovative Ni-IMAC Mini Kits combine the quality separation you expect
from gravity flow columns with the speed and ease-of-use of spin columns.
For
FPLC applications Super Ni-NTA and Co-NTA Agarose Resins are ideal.
SERVA's
standard und Superflow agarose resins in various formats (loose, pre-packed
columns, kits) and magnetic agarose beads allow protein purification from lab
up to process scale.