PerfectPro Ni-NTA MagBeads, 5PRIME

2400600
10052-316EA 257.46 CAD
10052-316
PerfectPro Ni-NTA MagBeads, 5PRIME
  • Reproducible purification of 6xHis-tagged proteins also from uncleared lysates
  • Wide range of binding capacities by simply varying the amount of beads used per well
  • Extremely low non-specific binding of proteins to bead surface
  • Easy to handle and ready-to-use matrices for any scale of purification- native or denaturing
  • High throughput screening and versatile magnetocapture

Magnetic Ni-NTA resin for high-throughput, microscale purification of 6xHis-tagged proteins. PerfectPro Ni-NTA MagBeads can be used for capturing 6xHis-tagged proteins under either native or denaturing conditions. PerfectPro Ni-NTA MagBeads are supplied as a 5% (v/v) suspension with a binding capacity of 300 µg protein per ml of suspension for 6xHis-tagged dihydrofolate reductase (DHFR, approximately 12.5 nmol per ml, molecular weight: 24 kDa), which is ideal for high-throughput, microscale purification of 6xHis-tagged proteins. PerfectPro Ni-NTA MagBeads are agarose beads that contain magnetic particles. The surface of the magnetic particles is covalently bound to strongly metal-chelating nitrilotriacetic acid (NTA) groups to their surfaces. The average diameter of a bead is 50 µm and the diameters range from 20 to70 µm.

Magnetic Agarose Beads allows flexible choice of the amount of 6xHis-tagged protein captured to suit the particular assay. As the beads are precharged with nickel, they are ready to use for high-throughput assays and screening programs, as well as small-scale purification of 6xHis-tagged proteins. The beads can be used for under either native or denaturing conditions. A wide variety of magnetocapture assays involve interaction of immobilized, functionally active 6xHis-tagged proteins with interacting molecules, such as proteins or nucleic acids.

The PerfectPro Ni-NTA System provides a complete system for the expression, purification, detection, and assay of 6xHis-tagged proteins. Protein expression with the PerfectPro Ni-NTA System begins with constructing expression clones, followed by the expression of 6xHis-tagged proteins and purification on PerfectPro matrices. High-throughput, micro-scale purification applications. Binding capacity: 0.25–2 mg/mL suspension (5%). Cross-linked 3% agarose bead structure. Bead size 50 µm average (range: 20–70 µm). Form 5% suspension in 30% ethanol, precharged with Ni2+. PerfectPro Ni-NTA System provides a complete system for the expression, purification, detection, and assay of 6xHis-tagged proteins. Protein expression with the PerfectPro Ni-NTA System begins with constructing expression clones, followed by the expression of 6xHis-tagged proteins and purification on PerfectPro matrices.

Purification of recombinant proteins using the PerfectPro Ni-NTA System does not depend on the 3-dimensional structure of the protein or 6xHis tag. This allows one-step protein purification under either native or denaturing conditions, from dilute solutions and crude lysates. Strong denaturants and detergents can be used for efficient solubilization and purification of receptors, membrane proteins, and proteins that form inclusion bodies. Reagents that allow efficient removal of nonspecifically binding contaminants can be included in wash buffers. Purified proteins are eluted under mild conditions by adding 100–250 mM imidazole as competitor or by a reduction in pH.

Ni-NTA MagBeads are very robust and is compatible with mostreagents usedfor protein purification. β-mercaptoethanol-Prevents disulfide cross-linkagesCan reduce nickel ions at higher concentration, up to 20 mM
DTT, DTE-Low concentrations will reduce nickel ions, a maximum of 1 mM may be used, but β-mercaptoethanol is recommended. Nonionic detergents (Triton, Tween, NP-40, etc.)-Removes background proteins and nucleic acids, up to 2% can be used
CHAPS-Up to 1% can be used. GuHCl-Solubilize proteins, up to 6 M. Urea-Solubilize proteins, up to 8 M. NaCl-Prevents ionic interactions, up to 2 M can be used, at least 300 mM should be used. MgCl2-Up to 4 M. CaCl2-Up to 5 mM. Glycerol-Prevents hydrophobic interaction between proteins, up to 50%. Ethanol-Prevents hydrophobic interactions between proteins, up to 20%. Imidazole-Binds to PerfectPro and competes with histidine residues in the 6xHis tag. Can be used at low concentrations (20 mM) to inhibit non-specific binding and, at higher concentrations to elute the 6xHis-tagged protein from the PerfectPro matrix.
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