GE Healthcare
Instructions 71-5017-51 AG HiTrap ion exchange columns HiTrap DEAE FF,1 ml and 5 ml HiTrap CM FF, 1 ml and 5 ml HiTrap Q FF, 1 ml and 5 ml
HiTrap SP FF, 1 ml and 5 ml HiTrap ANX FF (high sub), 1 ml and 5 ml
HiTrap™ SP FF, HiTrap CM FF, HiTrap Q FF, HiTrap DEAE FF, and HiTrap ANX FF (high sub) are prepacked 1 ml and 5 ml, ready to u cation and anion exchange HiTrap columns for method scouting, group parations, sample concentration and sample clean-up of charged biomolecules. The special design of the column, together with a modern chromatography media, provides fast, reproducible, and easy parations in a convenient format.
realityThe columns can be operated with a syringe, peristaltic pump or liquid chromatography system such as ÄKTA™ design.
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1Union 1/16” female/M6 male is also needed.2
Union M6 female/1/16” male is also needed.
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Code No. Product Type of Medium No. supplied
17-5157-01 HiTrap SP FF Strong cation exchanger 5 × 5 ml 17-5056-01 HiTrap CM FF Weak cation exchanger 5 × 1 ml 17-5155-01 HiTrap CM FF Weak cation exchanger 5 × 5 ml 17-5053-01 HiTrap Q FF Strong anion exchanger 5 × 1 ml 17-5156-01 HiTrap Q FF Strong anion exchanger 5 × 5 ml 17-5055-01 HiTrap DEAE FF Weak anion exchanger 5 × 1 ml 17-5154-01 HiTrap DEAE FFx
Weak anion exchanger
5 × 5 ml 17-5162-01 HiTrap ANX FF (high sub) Weak anion exchanger 5 × 1 ml 17-5163-01 HiTrap ANX FF (high sub) Weak anion exchanger
5 × 5 ml
Connectors supplied
Usage
No. supplied
1/16” male/luer female Connection of syringe to top of
HiTrap column 1Tubing connector flangeless/M6 female Connection of tubing (e.g. Peristaltic Pump P1) to bottom of HiTrap column 1 1Tubing connector flangeless/M6 male Connection of tubing (e.g. Peristaltic Pump P1) to top of HiTrap column 2 1Union 1/16” female/M6 male Connection to original FPLC™ System through bottom of HiTrap column 1Union M6 female/1/16” male
Connection to original FPLC System through top of HiTrap column
1Stop plug female, 1/16” Sealing bottom of HiTrap column
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Table of contents
1.Description (4)
2.General considerations (7)
3.Operation (13)
4.Purification (13)
5.Optimizing starting conditions (15)
会议桥6.Further optimization (16)
7.Determination of binding capacity (19)
8.Scaling up (20)
9.Storage (20)
10.Intended u (22)
11.Ordering information (22)
1Description
Column
The columns are 1 ml and 5 ml columns made of polypropylene, which is biocompatible and non-inte
ractive with biomolecules. The top and bottom frits are manufactured from porous polyethylene. It is delivered with a stopper on the inlet and snap-off end on the outlet.
The paration can be easily achieved using a syringe together with the supplied luer connector, a peristaltic pump, or in a chromatography system such as ÄKTA design.The column cannot be opened or refilled.
Note:
To prevent leakage it is esntial to ensure that the connector is tight.
Characteristics of HiTrap column are listed in Table 1.
Table 1. Characteristics of HiTrap column.Media Properties
Q Sepharo™ Fast Flow, DEAE Sepharo Fast Flow, SP Sepharo Fast Flow, and CM Sepharo Fast Flow are bad on a robust, 6% highly cross-linked beaded agaro matrix with excellent flow properties and high loading capacities.
产地英文ANX Sepharo 4 Fast Flow (high sub) is bad on 4% highly cross-linked beaded agaro. This re
sults in a medium with higher porosity, which is particularly uful for the purification of high molecular mass proteins.
Characteristics of the ion exchangers are listed in Table 2.
Column volumes 1 ml or 5 ml
Column dimensions 0.7 × 2.5 cm (1 ml) or 1.6 × 2.5 cm (5 ml)Maximum flow rates HiTrap 1 ml: 4 ml/min, HiTrap 5 ml: 20 ml/min Recommended flow rates
HiTrap 1 ml: 1ml/min, HiTrap 5 ml: 5 ml/min
Maximum back pressure 0.3 MPa, 3 bar
Chemical stability
All commonly ud aqueous buffers, 1 M
NaOH, 8 M urea, 6 M guanidine hydrochloride, 70% ethanol
Table 2. Characteristics of Sepharo Fast Flow ion exchangers
Cation exchangers SP Sepharo Fast Flow CM Sepharo Fast Flow
Bead structure 6% highly cross-linked
beir
agaro 6% highly cross-linked agaro
Bead size 45–165 µm 45–165 µm Type of gel Strong cation Weak cation Charged group -CH2CH2CH2SO3- -O-CH2COO-
Total ionic capacity 0.18–0.25 mmol H+/
ml medium 0.09–0.13 mmol H+/ ml medium
Dynamic binding capacity170 mg Ribonuclea A/
ml medium
50 mg Ribonuclea A/
ml medium
pH stability:
Short term23–14 2–14
Working4–13 6–10
Long term34–13 4–13
Storage
temperature
4°C to 30°C 4°C to 30°C
Storage buffer 20% ethanol,
cruelly0.2 M sodium acetate
20 % ethanol
Chemical stability All commonly ud aqueous buffers, 1 M NaOH, 8 M urea,
6 M guanidine hydrochloride, 70% ethanol
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memoAvoid Oxidizing agents, cationic detergents and buffers.
See Table3 and Figure 2.
