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HisTrap HP columns are prepacked with Ni Sepharose High Performance and designed for simple, hig↓h-resolution purification of histidine-tagged proteins by immobilized metal ion affinity chr≈omatography (IMAC).

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

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

  • Negligible Ni2+ leakage

  • Simple manual operation with a syringe, pump, or chromatography s©ystem 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 bβinding 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 conta✘ining shorter or longer polyhistidine tags, such as✘ (histidine)4 or (histidine)10. The shorter (histidine)4 will bind m✘ore weakly and the longer (histidine)10 will bind more strongly÷ compared with (histidine)6. This difference in binding str✔ength can be used during purification; since (histidi"ne)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 Per£formance maintains biological activity and increases product yield, at the same t♦ime as it greatly expands the range of suitable operating conditions.

For convenient scaling up of histidine-tagged protein purification, use 20 ml Hi↑sPrep 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 βof 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. Te sted 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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