His-tag Purification

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.

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).

GoldBio’s new Highest Density Nickel Agarose is a high capacity resin manufactured with a polychelator ligand. The product was developed to work in the presence of EDTA and DTT without any loss of performance. Its specificity and stability allows a one-step purification, eliminating the need for pretreatment of samples that cause nickel stripping.

Our Highest Density Nickel Agarose is supplied as a suspension in 50% aqueous suspension containing 20% ethanol. Purifications can be performed using 20mM EDTA and 20mM DTT and can work in both native as well as denaturing conditions.

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.

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.