Affinity Reagents

This matrix is a highly cross-linked 4% agarose bead with a bead size of 50-150 µm. It is a gel filtration resin developed for industrial-scale separation of large molecules and virus particles utilizing higher pressure and flow rates rather than low pressure standard beads.

This matrix is not preactivated. For preactivated resins, look for Glyoxal Agarose Beads or Protein G.

This Buffer Set consists of:

- 5X Phosphate Buffer, pH8 (250 mM Phosphate buffer pH 8, 1.5 M NaCl)
- 25 mL 3M Recrystallized Imidazole Solution, pH 6.0

This Buffer set allows researchers to develop, wash and elute buffers for His-tag protein purification.

5X Phosphate Buffer, pH8 (250 mM Phosphate buffer pH 8, 1.5 M NaCl).

Phosphate Buffer, pH8 is a mix of Sodium Phosphate dibasic (Na2HPO4) and Monosodium Phosphate (NaH2PO4). This buffer is the basis for wash and elution buffers for protein purification through His-tag.

This matrix is a highly crosslinked 6% agarose bead with a bead size of 50-150 µm. It is a gel filtration resin developed for industrial-scale separation of large molecules and virus particles utilizing higher pressure and flow rates rather than low pressure Standard Beads.

This matrix is not preactivated. For preactivated resins, look for Glyoxal Agarose Beads or Protein G.

Available in bulk or in prepacked columns, cobalt has a binding capacity of >25 mg/ml with approximately 60 kDa protein. Cobalt binds fewer host protein contaminants, resulting in lower background than nickel resins and no metal contamination in eluted histidine-tagged protein sample. In addition, it allows for purification of proteins under native or denaturing conditions. Cobalt Agarose works very well with Ni/Cu/Co Resin Wash Buffer and Histidine Protein Elution Buffer.

Cobalt HTC 6% Agarose Beads are suitable for downstream purification using IMAC. These resins combine the advantages of Cobalt purification with the high flow rates of the HTC resin (70% higher than commercial standards).

Glutathione has been covalently linked for use in affinity purification of glutathione-S-transferase (GST) and GST fusion proteins.

Biomolecules can be immobilized by binding them to a support under conditions that will then determine the characteristics of the complex. Immobilization is a technique that binds a ligand (enzyme, antibody, affinity proteins like Protein A or Protein G) to a support, giving high stability and making it easier to reuse the immobilized molecule.

Glyoxal beads allow a covalent binding of agarose to lysine amino groups (similar orientation to CNBr). Glyoxal agarose beads are ready to use and require no activation. The resin has a high amount of aldehydes groups (40-60 µmoles Glyoxal / ml gel) providing a very good option to conjugate affinity ligands improving the stability of the ligand.

The choice of Glyoxal or Aminoethyl will depend on the biomolecule to be immobilized, the accessibility of the reactive groups, and the direction/ orientation required for the binding to the support.

Biomolecules can be immobilized by binding them to a support under conditions that will then determine the characteristics of the complex. Immobilization is a technique that binds a ligand (enzyme, antibody, affinity proteins like Protein A or Protein G) to a support, giving high stability and making it easier to reuse the immobilized molecule.

Glyoxal beads allow a covalent binding of agarose to lysine amino groups (similar orientation to CNBr). Glyoxal agarose beads are ready to use and require no activation. The resin has a high amount of aldehydes groups (15-25 µmoles Glyoxal / ml gel) providing a very good option to conjugate affinity ligands improving the stability of the ligand.

The choice of Glyoxal or Aminoethyl will depend on the biomolecule to be immobilized, the accessibility of the reactive groups, and the direction/ orientation required for the binding to the support.

Glyoxal HTC 6% HIGH Density Agarose beads allow covalent binding of agarose to lysine amino groups (similar orientation to CNBr).

This product is adequate to work in large scale process (High Pressure). The resin has a high amount of aldehyde groups (40-60 µmoles Glyoxal / ml gel) providing multiple binding points and high immobilized ligand stability. The resin has been manufactured using 6% HTC agarose beads, and the pore size is adequate to work with enzymes, proteins & small ligands. The stability at high flow is adequate for large scale process.

His-Tag Buffer Set includes:

- 250 mL Wash Buffer (50mM Phosphate Buffer pH 8, 300mM NaCl, 10mM Imidazole)
- 50 mL Elution Buffer (50mM Phosphate Buffer, 300mM NaCl, 500mM Imidazole)

Designed for use with Goldbio Nickel and Cobalt Agarose resins, but it can be used with most standard His-tag purification resins.

Imidazole is a planar 5-membered ring. It is a highly polar, highly water soluble amphoteric compound. Imidazole is often used to elute tagged proteins bound to metal ions attached to the surface of beads in a chromatography column by displacing the His-tag from the metal ion and freeing the His-tagged proteins.

Imidazole is a planar 5-membered ring. It is a highly polar, highly water soluble amphoteric compound. Imidazole is often used to elute tagged proteins bound to metal ions attached to the surface of beads in a chromatography column by displacing the His-tag from the metal ion and freeing the His-tagged proteins.

Pepstatin A is a competitive, reversible aspartic protease inhibitor which inhibits proteases such as pepsin, chymosin, renin, HIV proteases, and cathepsins D and E. Peptstatin A is highly selective and is often used in a mixture with other enzyme inhibitors. In vitro studies have shown pepstatin A is the only aspartic acid inhibitor that exhibits inhibitory effects on HIV proteases.

Nickel Agarose Resin is available in bulk or in prepacked columns. It has a binding capacity of >25 mg/ml with approximately 60 kDa protein. In addition, it allows for purification of proteins under native or denaturing conditions. GoldBio Nickel Agarose works very well with the His-Tag Buffer Set.

IDA cross-linked Agarose resin consists of iminodiacetic acid groups ligated by stable ether linkages via a spacer arm. IDA is a tridentate chelating agent, covalently coupled to cross-linked agarose beads. This resin is loaded with Ni2+. The resulting, ready-to-use resin is ideal for rapid purifications of His-tagged proteins.

Affinity Chromatography is based on the interaction between certain superficial protein residues (histidines, cysteines and to a lesser extent tryptophans), with transition metal cations, forming chelates.

Low Density Nickel Agarose Resin is available in bulk or in prepacked columns. It has a binding capacity of >12 mg/ml with approximately 60 kDa protein. It works very well with the His-Tag Buffer Set.

IDA cross-linked Agarose resin consists of iminodiacetic acid groups ligated by stable ether linkages via a spacer arm. IDA is a tridentate chelating agent, covalently coupled to cross-linked agarose beads. This resin is loaded with Ni2+. The resulting, ready-to-use resin is ideal for rapid purifications of His-tagged proteins.

Affinity Chromatography is based on the interaction between certain superficial protein residues (histidines, cysteines and to a lesser extent tryptophans), with transition metal cations, forming chelates.

Nickel HTC 6% Agarose Beads are suitable for downstream purification using IMAC. These resins combine the advantages of Nickel purification with the high flow rates of the HTC resin (70% higher than commercial standards).

HTC beads are high throughput beads designed to meet the demand for industrial process separation. Their rigidity and mechanical resistance permits high flow rates, with good resolution in a minimum time frame, making these beads ideal for process-scale use. HTC beads are highly crosslinked 4% and 6% agarose matrices and have excellent physical and chromatographic qualities. They are autoclavable. These beads are an ideal support for the immobilization of ligands for Affinity Chromatography and base media support for producing IEX (Ion Exchange) and Hydrophobic interaction chromatography resins.

Nickel-NTA Agarose Resin has a binding capacity of ~50 mg/ml. In addition, it allows for purification of proteins under native or denaturing conditions. GoldBio Nickel Agarose works very well with the His-Tag Buffer Set.

NTA cross-linked Agarose resin consists of nitrilotriacetic acid groups ligated by stable ether linkages via a spacer arm. NTA is a tetravalent chelating agent, covalently coupled to cross-linked agarose beads, providing a higher specificity and lower ion leaching than IDA linked resins. NTA resins have also been shown to be more robust in the presence of higher concentrations of EDTA, but may require a higher imidazole concentration for protein elution. This resin is loaded with Ni 2+. The resulting, ready-to-use resin is ideal for rapid purifications of His-tagged proteins.

Nickel NTA HTC Agarose Resin has a binding capacity of ~60 mg/ml. In addition, it allows for purification of proteins under native or denaturing conditions. GoldBio Nickel Agarose works very well with the His-Tag Buffer Set.

NTA cross-linked Agarose resin consists of nitrilotriacetic acid groups ligated by stable ether linkages via a spacer arm. NTA is a tetravalent chelating agent, covalently coupled to cross-linked agarose beads, providing a higher specificity and lower ion leaching than IDA linked resins. NTA resins have also been shown to be more robust in the presence of higher concentrations of EDTA, but may require a higher imidazole concentration for protein elution. This resin is loaded with Ni 2+. The resulting, ready-to-use resin is ideal for rapid purifications of His-tagged proteins.

Nickel NTA Magnetic Agarose Beads are a rapid and easy small scale purification of histidine-tagged proteins. This resin consists of magnetic agarose derivatized with Nitrilotriacetic (NTA) and provides good properties working in native or denaturing conditions. Magnetic agarose beads provide a convenient and quick method for purification without the need for pipetting or centrifugation. Washing, binding and elution steps will require a magnetic device.

NTA cross-linked resins consist of nitrilotriacetic acid groups ligated by stable ether linkages via a spacer arm. NTA is a tetravalent chelating agent which provides a higher specificity and lower ion leaching than IDA linked resins. NTA resins have also been shown to be more robust in the presence of higher concentrations of EDTA, but may require a higher imidazole concentration for protein elution. This resin is loaded with Ni 2+. The resulting, ready-to-use resin is ideal for rapid purifications of His-tagged proteins.