High capacity, single-step purification of His tag proteins. Ready to use columns for AKTA and other automated systems. ~40 mg His tagged protein/mL medium. ... Read more
PROTEINDEX™ HiBond™ Ni-NTA Agarose 6FF has been optimized for automated purification applications. It has been widely used in scale-up and preparative purification of histidine-tagged proteins that are expressed in various hosts, such as E.coli, yeast, insect cells, and mammalian cells.
The base matrix of PROTEINDEX™ HiBond™ Ni-NTA Agarose 6FF is highly crosslinked 6% agarose, offering excellent chemical-, and mechanical stability for automated applications. The matrix support is functionalized with the chelate ligand Nitilotriacetic Acid (NTA) in complex with the nickel ions (Ni2+) in a very stable octahedral structure. The structure of Ni-NTA complex is compatible solutions that contain certain concentration of reducing agents, denaturing agents, detergents and other additives.
The NTA ligand coordinates the Ni2+ with four valencies (tetradentate, coordination number 4), and two valencies are available for interaction with imidazole rings of histidine residues. The number of available valencies seems to provide the best combination of binding effectiveness and chemical stability for binding with the His tag. The Ni-NTA ion-chelate complex on our ProteIndex™ HiBond™ Ni-NTA resins has been delicately optimized to ensure the most effective capture for His tag purification of His-tagged proteins.The Prepacked Cartridge has standard interface and can be adapted to most commonly used chromatography systems, such as ÄKTA systems.
- Convenient, easy, ready-to-use
- Mechanical stable, designed for large scale purification of histidine-tagged proteins
- High yield (typically > 40 mg protein /mL resins)
- Low leakage of nickel ions. Can be re-used for up to 8 preps*
- Automated purifications
- Routine His tag protein purification for structural biology and functional assays
- Scaling up and preparative applications
One-step purification using ProteIndex™ HiBond™ Ni-NTA Agarose 6 Fast Flow resins
Figure 2. A 25 kDa protein was expressed in E.coli and purified on an automated system using a one-step automated protocol with ProteIndex™ HiBond™ Ni-NTA Agarose 6 Fast Flow resin under native conditions. 50 mL E.coli lysate in 50 mM NaH2PO4, 0.3 M NaCl, 10 mM imidazole was loaded on the column packed with 0.5 mL resins at the speed of 1 mL/min. The column was then washed with 100 mL 50 mM NaH2PO4, 0.3 M NaCl, 20 mM imidazole at the speed of 1~2 mL/min. The target protein was eluted with 50 mM NaH2PO4, 0.3 M NaCl, 250 mM imidazole. A fraction of eluate was analyzed on a SDS-PAGE. Shown were coomassie blue stains of the target proteins purified from columns of Marvelgent’s ProteIndex Ni-NTA 6FF (M) and Ni-Sepharose (G) resins.
|Binding capacity||>40 mg 6xHis-tagged protein/mL medium|
|Matrix||6% highly cross-linked agarose|
|Particle size||45 μm - 165 μm|
|Column volume||1 mL or 5 mL|
||0.3 MPa (3 bar)|
|Column hardware pressure limit||5 bar (0.5 MPa)|
||2 - 8°C|
||1x PBS containing 20% ethanol|
Product is shipped at ambient temperature. Upon receipt store at 2 - 8°C. Do not Freeze.
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