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In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
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HyperChrom Ni-NTA Excel Agarose is an affinity chromatography medium that chemically couples a tetracoordinated transition metal ion Ni2+ to a ligand of Nitrilotriacetic acid (NTA) using a highly cross-linked agarose gel as a matrix. It has a very stable octahedral structure, with nickel ions in the center of the octahedron to protect nickel ions from small molecules, is more stable, and can withstand harsh conditions such as reducing agents, denaturants or couplants at certain concentrations. In addition, due to the pressure resistance of the matrix, which can withstand pressures up to 0.3 MPa, the product can be used for industrial-scale protein purification, enabling purification of the protein of interest at relatively high flow rates. This chromatography medium uses the interaction between Ni2+ and the side chains of certain amino acids (mainly histidine) on the protein to separate and purify proteins with and without these amino acids, and with or without these amino acids.
HyperTrap Ni-NTA Excel ColumnReady-to-use columns with HyperChrom Ni-NTA Excel Agarose can be used to purify histidine-tagged proteins with tolerances such as 100 mM EDTA and 0.5 M NaOH immersion for 48 h.
1. HyperTrap Ni-NTA Excel Column preloaded column parameters
Prepacked column specifications/names
1 mL
5 mL
Dimensions (IDxH)
7.7 x 25 mm
16 x 25 mm
Pressure-resistant
0.3 MPa
Workflow speed*
1 mL/min
1-3 mL/min
Operating temperature
4 - 30℃
Storage conditions
Store the components at 4℃ for 5 years.
*This flow rate is the recommended common flow rate, the pressure resistance, and the flow rate of different types of chromatography resins in the prepacked column will be different, please refer to the product manual of the corresponding chromatography resin.
2. HyperTrap Ni-NTA Excel Column chromatography media parameters
Name
Description
Chromatography media type
Affinity chromatography media
Ligands
NTA-Ni2+
Scaffolding
Highly cross-linked agarose
Average particle size
90 μm
Ligand density
54~70 μmol Ni2+/mL chromatography medium
Dynamic load
≥ 10 mg histidine-tagged protein/mL chromatography medium
Flow rate is recommended
150-600 cm/h
Maximum flow rate
600 cm/h
pH stability *
2-14
Chemical resistance
0.1 M~0.5 M NaOH for 48 h
10 mM β-mercaptoethanol, 5 mM TCEP for 24 h
500 mM imidazole, 100 mM EDTA, 2 h
*After 7 days of chromatography media at 40°C, pH 2-14, there was no significant change in its physicochemical properties and functions.