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In vitro transcription of capped mRNA with modified nucleotides and Poly(A) tail
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HpyerChrom Ni-NTA HP Agarose is an affinity chromatography medium made by chemically coupling the transition metal ion Ni2+ with tetracoordination to a ligand of Nitrilotriacetic acid (NTA) using a highly crosslinked agarose gel as a matrix. It has a very stable octahedral structure, nickel ions are at the center of the octahedron, which protects nickel ions from attack by small molecules, is more stable, and can withstand harsh conditions such as reducing agents, denaturants or couplants in a certain concentration. 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 large-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 achieve the separation and purification of proteins containing and not containing these amino acids and containing a large and low number of these amino acids.
HyperTrap Ni-NTA HP Column, a ready-to-use chromatography column with HyperChrom Ni-NTA HP Agarose as the filler, can be used to purify histidine-tagged proteins, which has the advantages of high performance, high load, easy regeneration and low cost.
1. HyperTrap Ni-NTA HP Column preloaded column parameters
Pre-assembled column specification/name
1 mL
5 mL
Dimensions (IDxH)
7.7 x 25 mm
16 x 25 mm
Withstand pressure
0.3 MPa
Workflow Speed*
1 mL/min
1-3 mL/min
Operating temperature
4 - 30°C
Storage condition
Store the components at 4℃ for 5 years.
*This flow rate is the recommended common flow rate, the pressure resistance, and the use of different types of chromatography packing in the pre-assembled column will be different, please refer to the product manual of the corresponding chromatographic filler.
2. HyperChrom Ni-NTA HP Agarose chromatography media parameters
Name
Description
Chromatography media type
Affinity chromatography media
Ligation
NTA-Ni2+
Scaffolding
Highly cross-linked agarose
Average particle size
34 μm
Ligand density
~15 μmol Ni2+/mL chromatography medium
Dynamic load
≥40 mg histidine-tagged protein/mL chromatography medium*
Flow rates are recommended
90-150 cm/h
Maximum flow rate
200 cm/h
Use temperature
4 - 30℃
pH stability **
2-14
Chemical resistance
Common aqueous solution, 0.01 M HCl, 0.1 M NaOH, 8 M urea, 6 M guanidine hydrochloride, 30% isopropanol ***
* Dynamic load measurement conditions: column loading height: 10 cm, test flow rate 150 cm/h; Test buffer: 0.02 M PB, 0.5 M NaCl, 5 mM imidazole, pH 7.4; test sample: tagged protein (1 mg/ml, 6*His); Index: When the protein penetration reaches 10%, the amount of protein loaded per unit volume of media (mL) (mg).
**After 7 days in the environment of chromatography at 40 °C and pH 2-14, the physical and chemical properties and functions of the chromatography medium did not change significantly.
*** 30% is v/v, volume ratio.