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HisTrap HP columns are prepacked with Ni Sepharose High Performance and designed for simple, high-resolution purification of histidine-tagged proteins by immobilized metal ion affinity chromatography (IMAC).

  • High binding capacity, at least 40 mg/ml chromatography medium.

  • Compatible with a wide range of reducing agents, detergents, denaturants, and other additives.

  • Negligible Ni2+ leakage

  • Simple manual operation with a syringe, pump, or chromatography system such as ÄKTA design

HisTrap HP 1 ml and 5 ml columns are designed for simple, one-step purification of histidine-tagged proteins. The columns are prepacked with Ni Sepharose High Performance, which has high binding capacity and low nickel ion leakage that ensures reliable capture of target protein in repeated IMAC purifications.

HisTrap HP columns can also be used for the purification of tagged proteins containing shorter or longer polyhistidine tags, such as (histidine)4 or (histidine)10. The shorter (histidine)4 will bind more weakly and the longer (histidine)10 will bind more strongly compared with (histidine)6. This difference in binding strength can be used during purification; since (histidine)10 binds more strongly, a higher concentration of imidazole can be added to the lysed cells. This can facilitate the removal of contaminants that can otherwise be copurified with the tagged target protein.

The high stability and broad compatibility of Ni Sepharose High Performance maintains biological activity and increases product yield, at the same time as it greatly expands the range of suitable operating conditions.

For convenient scaling up of histidine-tagged protein purification, use 20 ml HisPrep FF 16/10 columns. Ni Sepharose 6 Fast Flow, the medium prepacked in HisTrap FF and HisPrep FF 16/10 columns, allows high flow rates, which facilitates scale-up of histidine-tagged protein purification.

Bed Dimensions16 mm × N/A
Bed Volume5 ml
Binding Capacity/ColumnAt least 40 mg histidine-tagged protein/ml medium1)
Flow Rate4 ml/min (1 ml), 5 ml/min (5ml)2)
Storage Conditions4 to 30°C, 20% Ethanol
Pressure Max. [Over the Packed Bed During Operation]N/A (N/A)
1)Protein binding capacity is protein-to-protein dependent.
2)H₂0 at room temperature


Media
Particle size, d50V34 µm1)
MatrixHighly cross-linked agarose, 6%
Binding Capacity/ml Chromatography MediumAt least 40 mg histidine-tagged protein/ml medium2)
Metal Ion Capacityapprox. 15 µmol Ni2+/ml medium
pH Stability Working Range3–123)
pH Stability Cleaning2–144)
pH stability, CIP2–145)
Storage Conditions4 to 30°C, 20% Ethanol
Chemical StabilityFor Ni2+-stripped medium 0.01 M HCl, 0.1 M NaOH. Tested for one week at 40°C 1 M NaOH, 70% HAc. Tested for 12 h 2% SDS. Tested for 1 h 30% 2-propanol. Tested for 30 min
Avoid UsingChelating agents such as EDTA, EGTA , citrate
1)d50v is the median particle size of the cumulative volume distribution.
2)Protein binding capacity is protein-to-protein dependent.
3)Ni²?-stripped medium.
4)Ni²?-stripped medium.
5)Ni²?-stripped medium.


Column
Bed Dimensions16 mm × N/A
Column i.d.16 mm
Complete Packsize5 ml
Material [Column Hardware]Polypropylene (PP)

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