HisTrap+HP,+1+ml+and+5+ml
GE Healthcare
Instructions 71-5027-68 AH HisTrap affinity columns HisTrap HP, 1 ml and 5 ml
HisTrap? HP is a ready to use column, prepacked with precharged Ni Sepharose? High Performance. This prepacked column is ideal for preparative purification of Histidine-tagged recombinant proteins by immobilized metal ion affinity chromatography (IMAC).
The special design of the column, together with the high-performance matrix of the Ni Sepharose medium, provides fast, simple, and easy separations in a convenient format.
Ni Sepharose High Performance has low nickel (Ni2+) ion leakage and is compatible with a wide range of additives used in protein purification. HP columns can be operated with a syringe, peristaltic pump, or liquid chromatography system such as ?KTA? design chromatography systems.
CAUTION! Contains nickel. May produce an allergic reaction.
.
1
Pack size available by special order.
Connector kit
1Union 1/16” female/M6 male is also needed.
2
Union M6 female/1/16” male is also needed.
Table of contents
1.Description..............................................................................32.General considerations.....................................................63.Operation.................................................................................74.Optimization........................................................................115.Stripping and recharging..............................................126.Cleaning-in-place.............................................................137.Scaling-up............................................................................138.Storage..................................................................................149.Troubleshooting................................................................1410.Intended use.......................................................................1711.Ordering Information (18)
Code No. Product No. supplied 17-5247-01 HisTrap HP 5 × 1 ml 17-5247-05 HisTrap HP 100 × 1 ml 117-5248-01 HisTrap HP 1 × 5 ml 17-5248-02 HisTrap HP 5 × 5 ml 17-5248-05
HisTrap HP 100 × 5 ml 1
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 2, 5 or 7
1Description
Medium properties
HisTrap HP 1 ml and 5 ml columns are prepacked with Ni
Sepharose High Performance, which consists of 34 μm highly
cross-linked agarose beads with an immobilized chelating group.
The medium has then been charged with Ni2+-ions.
Several amino acids, for example histidine, form complexes with many metal ions. Ni Sepharose High Performance selectively binds proteins if suitable complex-forming amino acid residues are
exposed on the protein surface.
Additional histidines, such as in the case of (histidine)6 -tag,
increase affinity for Ni2+ and generally make the histidine-tagged protein the strongest binder among other proteins in for example an E. coli extract.
Column properties
HisTrap HP columns are made of biocompatible polypropylene
that does not interact with biomolecules. Columns are delivered
with a stopper on the inlet and a snap-off end on the outlet. The
columns have porous top and bottom frits that allow high flow
rates. They cannot be opened or refilled.
Columns can be operated with either a syringe and the supplied luer connector, a peristaltic pump, or a chromatography system
such as ?KTA design.
Note:To prevent leakage, ensure that the connector is tight.
Table 1. HisTrap HP characteristics.1
Dynamic binding capacity conditions:Note: Dynamic binding capacity is protein-dependent.2H 2O at room temperature 3
Ni 2+-stripped medium
Matrix
Highly cross-linked spherical agarose, 6%Average particle size 34 μm
Metal ion capacity ~ 15 μmol Ni 2+/ml medium
Dynamic binding capacity 1 At least 40 mg (histidine)6-tagged protein/ml medium Column volumes 1 ml or 5 ml Column dimensions i.d. × H:0.7 × 2.5 cm (1 ml) 1.6 × 2.5 cm (5 ml)
Recommended flow rate 1 and 5 ml/min for 1 and 5 ml column respectively
Max. flow rates 4 and 20 ml/min for 1 and 5 ml column respectively Max back pressure 2
0.3 MPa, 3 bar
Compatibility during use Stable in all commonly used buffers, reducing
agents, denaturants, and detergents (s ee Table 2)Chemical stability 3
0.01 M HCl, 0.1 M NaOH. Tested for 1 week at 40°C.1 M NaOH, 70% acetic acid. Tested for 12 hours.2% SDS. Tested for 1 hour.
30% 2-propanol. Tested for 30 min.Avoid in buffers Chelating agents, e.g. EDTA, EGTA, citrate (see Table 2)
pH stability 3
short term (at least 2 hours): 2 to 14 long term (≤ 1 week): 3 to 12Storage 20% ethanol Storage temperature
4°C to 30°C
Sample:
1 mg/ml (histidine)6-tagged pure protein (M r 28 000 or 43 000) in
binding buffer (Q B, 10% determination) or (histidine)6-tagged protein bound from E. coli extract
Column volume: 0.25 ml or 1 ml Flow rate: 0.25 ml/min or 1 ml/min Binding buffer: 20 mM sodium phosphate, 0.5 M NaCl, 5 mM imidazole, pH 7.4Elution buffer: 20 mM sodium phosphate, 0.5 M NaCl, 0.5 M imidazole, pH 7.4
The Ni 2+-charged medium is compatible with all commonly used aqueous buffers, reducing agents, denaturants such as 6 M Gua-HCl and 8 M urea, and a range of other additives (see Table 2).
Table 2. Ni Sepharose High Performance is compatible with the following compounds, at least at the concentrations given.1See Notes and blank run, p. 10–11.2Tested for 1 week at 40°C.
3
The strong chelator EDTA has been used successfully in some cases, at 1 mM.
Generally, chelating agents should be used with caution (and only in the sample, not the buffers). Any metal-ion stripping may be counteracted by addition of a small excess of MgCl 2 before centrifugation/filtration of the sample. Note that stripping effects may vary with applied sample volume.
Reducing agents 1
5 mM DTE 5 mM DTT
20 mM ?-mercaptoethanol 5 mM TCEP
10 mM reduced glutathione Denaturing agents 8 M urea 2
6 M guanidine hydrochloride 2Detergents
2% Triton? X-100 (nonionic)2% Tween? 20 (nonionic)2% NP-40 (nonionic)2% cholate (anionic)1% CHAPS (zwitterionic)Other additives
500 mM imidazole 20% ethanol 50% glycerol 100 mM Na 2SO 41.5 M NaCl 1 mM EDTA 360 mM citrate 3
Buffer substances
50 mM sodium phosphate, pH 7.4100 mM Tris-HCl, pH 7.4100 mM Tris-acetate, pH 7.4100 mM HEPES, pH 7.4100 mM MOPS, pH 7.4
100 mM sodium acetate, pH 42
2General considerations
HisTrap HP is supplied precharged with Ni2+ ions. In general, Ni2+ is the preferred metal ion for purification of recombinant histidine-
tagged proteins. Note, however, that in some cases it may be wise to test other metal ions, for example Zn2+ and Co2+, as the strength of binding depends on the nature of the histidine-tagged protein as well as the metal ion (see Optimization).
We recommend binding at neutral to slightly alkaline pH (pH 7–8) in the presence of 0.5–1.0 M NaCl. Sodium phosphate buffers are often used. Tris-HCl can generally be used, but should be avoided in cases where the metal-protein affinity is very weak, since it may reduce binding strength. Avoid chelating agents such as EDTA or citrate in buffers, see Table2.
Including salt, for example 0.5–1.0 M NaCl in the buffers and
samples, eliminates ion-exchange effects but can also have a
marginal effect on the retention of proteins.
Imidazole at low concentrations is commonly used in the binding and the wash buffers to minimize binding of host cell proteins. For the same reason, it is important to also include imidazole in the
sample (generally, at the same concentration as in the wash
buffer). At somewhat higher concentrations, imidazole may also
decrease the binding of histidine-tagged proteins. The imidazole concentration must therefore be optimized to ensure the best
balance of high purity (low binding of host cell proteins) and high yield (binding of histidine-tagged target protein). This optimal
concentration is different for different histidine-tagged proteins,
and is usually slightly higher for Ni Sepharose High Performance than for similar IMAC media on the market (see Data File
18-1174-40). Use highly pure imidazole; such imidazole gives
essentially no absorbance at 280 nm.
As alternatives to imidazole elution, histidine-tagged proteins can be eluted from the medium by several other methods or
combinations of methods – a lowering of pH within the range of
2.5–7.5 can be used, for example. At pH values below 4, metal ions
will be stripped off the medium.
Note:If the proteins are sensitive to low pH, we recommend
collection of the eluted fractions in tubes containing
1 M Tris-HCl, pH 9.0 (60–200 μl/ml fraction) to restore the
pH to neutral.
Chelating agents such as EGTA or EDTA will also strip metal ions
from the medium and thereby cause protein elution, but the target protein pool will then contain Ni2+ ions. In this case, Ni2+ ions can be removed by desalting on a HiTrap? Desalting, a PD-10 Desalting Column, or HiPrep? 26/10 Desalting, (see Table3).
Leakage of Ni2+ from Ni Sepharose High Performance is very low under all normal conditions, lower than for other IMAC media
tested. For applications where extremely low leakage during
purification is critical, leakage can be even further reduced by
performing a blank run (see page 11). Likewise, a blank run should also be performed before applying buffers/ samples containing
reducing agents (see page 11).
Whatever conditions are chosen, HisTrap HP columns can be
operated with a syringe, peristaltic pump, or chromatography
system.
Note:If Peristaltic Pump P-1 is used, the maximum flow rate that can be run on a HisTrap HP 1 ml column is 3 ml/min.
3Operation
Buffer preparation
Water and chemicals used for buffer preparation should be of high purity. Filter buffers through a 0.22 μm or a 0.45 μm filter before
use.
Use high purity imidazole as this will give very low or no
absorbance at 280 nm.
If the recombinant histidine-tagged protein is expressed as
inclusion bodies, include 6 M Gua-HCl or 8 M urea in all buffers and sample. On-column refolding of the denatured protein may be
possible.
Recommended conditions
Sample preparation
For optimal growth, induction, and cell lysis conditions for your recombinant histidine-tagged clones, please refer to established protocols.
Adjust the sample to the composition and pH of the binding buffer by: Adding buffer, NaCl, imidazole, and additives from
concentrated stock solutions; by diluting the sample with binding buffer; or by buffer exchange, (see Table 3). Do not use strong bases or acids for pH-adjustment (precipitation risk). Filter the sample through a 0.22 μm or a 0.45 μm filter and/or centrifuge it immediately before applying it to the column.
To prevent the binding of host cell proteins with exposed histidine, it is essential to include imidazole at a low concentration in the sample and binding buffer (see Optimization).
Binding buffer: 20 mM sodium phosphate, 0.5 M NaCl,
20–40 mM imidazole, pH 7.4 (The optimal imidazole concentration is protein-dependent; 20–40 mM is suitable for many proteins.)Elution buffer:
20 mM sodium phosphate, 0.5 M NaCl, 500 mM imidazole, pH 7.4 (The imidazole concentration required for elution is protein-dependent).
Table 3. Prepacked columns for desalting and buffer exchange
Purification
1Fill the syringe or pump tubing with distilled water. Remove the stopper and connect the column to the syringe (use the luer
connector provided), laboratory pump or chromatography
system tubing “drop-to-drop” to avoid introducing air into the
system.
2Remove the snap-off end at the column outlet.
3Wash the column with 3–5 column volumes of distilled water.
4Equilibrate the column with at least 5 column volumes of binding buffer. Recommended flow rates are 1 ml/min or
5 ml/min for the 1 and 5 ml columns respectively.
In some cases a blank run is recommended before final
equilibration/ sample application (see page 11).
5Apply the pretreated sample using a syringe or a pump.
6Wash with binding buffer until the absorbance reaches a steady baseline (generally, at least 10–15 column volumes).
Note:Purification results are improved by using imidazole in
sample and binding buffer (see Optimization).
7Elute with elution buffer using a one-step or linear gradient.
Five column volumes are usually sufficient if the protein of
interest is eluted by a one-step gradient. A shallow gradient, for
example a linear gradient over 20 column volumes or more,
may separate proteins with similar binding strengths.
Note:If imidazole needs to be removed from the protein, use HiTrap Desalting, a PD-10 Desalting Column, or HiPrep
26/10 Desalting depending on the sample volume (see
Table3).
Note:Ni Sepharose High Performance is compatible with
reducing agents. However, removal of any weakly bound
Ni2+ ions by performing a blank run without reducing
agents (as described on page 11) before applying buffer/
sample including reducing agents is recommended. Do not
leave HisTrap HP columns with buffers including reducing
agents when not in use.
Note:Leakage of Ni2+ from Ni Sepharose High Performance is low under all normal conditions. The leakage is lower than for
other IMAC media tested (see Data File Ni Sepharose High
Performance, 18-1174-40). For very critical applications,
leakage during purification can be even further diminished
by performing a blank run (as described below) before
loading sample.
Blank run:
Use binding buffer and elution buffer without reducing agents.
1Wash the column with 5 column volumes of distilled water (to remove the 20% ethanol).
2Wash with 5 column volumes of elution buffer.
3Equilibrate with 10 column volumes of binding buffer.
4Optimization
Imidazole at low concentrations is commonly used in the binding and the wash buffers to minimize binding of host cell proteins. For the same reason, it is important to also include imidazole in the
sample (generally, at the same concentration as in the wash
buffer). At somewhat higher concentrations, imidazole may also
decrease the binding of histidine-tagged proteins. The imidazole concentration must therefore be optimized to ensure the best
balance of high purity (low binding of host cell proteins), and high yield (binding of histidine-tagged target protein). This optimal
concentration is different for different histidine-tagged proteins,
and is usually slightly higher for Ni Sepharose High Performance than for similar IMAC media on the market (see Data File Ni
Sepharose High Performance, 18-1174-40). Finding the optimal
imidazole concentration for a specific histidine-tagged protein is a trial-and-error effort, but 20–40 mM in the binding and wash
buffers is a good starting point for many proteins. Use a high purity imidazole, such imidazole gives essentially no absorbance at
280 nm.
Ni2+ is usually the first choice metal ion for purifying most
(histidine)6-tagged recombinant proteins from nontagged host cell proteins, and also the ion most generally used. Nevertheless, it is
not always possible to predict which metal ion will be best for a
given protein. The strength of binding between a protein and a
metal ion is affected by several factors, including the length,
position, and exposure of the affinity tag on the protein, the type of ion used, and the pH of buffers, so some proteins may be easier to purify with ions other than Ni2+.
A quick and efficient way to test this possibility and optimize
separation conditions is to use HiTrap IMAC HP 1 ml columns,
which are packed with IMAC Sepharose High Performance (not
charged with metal ions). Each column can be charged with
different metal ions, for example Cu2+, Co2+, Zn2+, Ca2+, or Fe2+.
Instructions are included with each column.
A study to compare the purification of six (histidine)6-tagged
recombinant proteins, including three variants of maltose-binding protein, with different metal ions has indicated that Ni2+ generally gives best selectivity between (histidine)-tagged and nontagged
host-cell proteins (see Application Note 18-1145-18).
5Stripping and recharging
Note:The column does not have to be stripped and recharged between each purification if the same protein is going to be
purified; it is sufficient to strip and recharge it after 5–7
purifications, depending on the cell extract, extract volume,
target protein, etc.
Recommended stripping buffer: 20 mM sodium phosphate,
0.5 M NaCl, 50 mM EDTA, pH 7.4
Strip the column by washing with at least 5–10 column volumes of stripping buffer. Wash with at least 5–10 column volumes of
binding buffer and 5–10 column volumes of distilled water before recharging the column.
Recharge the water-washed column by loading 0.5 ml or 2.5 ml of
0.1 M NiSO4 in distilled water on HisTrap HP 1 ml and 5 ml column,
respectively. Salts of other metals, chlorides, or sulfates, may also be used (see“Optimization”). Wash with 5 column volumes distilled water, and 5 column volumes binding buffer (to adjust pH) before storage in 20% ethanol.
6Cleaning-in-place
When an increase in back-pressure is seen, the column can be
cleaned. Before cleaning, strip off Ni2+ ions using the
recommended procedure described above.
After cleaning, store in 20% ethanol (wash with 5 column volumes) or recharge with Ni2+ prior to storage in ethanol.
The Ni2+-stripped column can be cleaned by the following
methods;
?Remove ionically bound proteins by washing the column with several column volumes of 1.5 M NaCl; then wash the column
with approximately 10 column volumes of distilled water.
?Remove precipitated proteins, hydrophobically bound proteins, and lipoproteins by washing the column with 1 M
NaOH, contact time usually 1–2 hours (12 hours or more for
endotoxin removal). Then wash the column with
approximately 10 column volumes of binding buffer, followed
by 5–10 column volumes of distilled water.
?Remove hydrophobically bound proteins, lipoproteins, and lipids by washing the column with 5–10 column volumes 30%
isopropanol for about 15–20 minutes. Then wash the column
with approximately 10 column volumes of distilled water.
Alternatively, wash the column with 2 column volumes of
detergent in a basic or acidic solution. Use, for example,
0.1–0.5% nonionic detergent in 0.1 M acetic acid, contact time
1–2 hours. After treatment, always remove residual detergent
by washing with at least 5 column volumes of 70% ethanol.
Then wash the column with approximately 10 column volumes
of distilled water.
7Scaling-up
Two or three HisTrap HP 1 ml or 5 ml columns can be connected in series for quick scale-up (note that back-pressure will increase).
Ni Sepharose High Performance, the medium prepacked in
HisTrap HP columns, is supplied preswollen in 25 and 100 ml lab
packs (see Ordering Information). An alternative scale-up strategy is thus to pack the medium in empty columns – Tricorn? and XK columns are suitable for this purpose.
8Storage
Store HisTrap HP columns in 20% ethanol at 4°C to 30°C.
9Troubleshooting
The following tips may be of assistance. If you have any further
questions about your HisTrap HP column, please visit
https://www.wendangku.net/doc/313264905.html,/hitrap, contact our technical support, or your local representative.
Note:When using high concentrations of urea or Gua-HCl,
protein unfolding generally takes place. Refolding on-
column (or after elution) is protein-dependent.
Tips:To minimize dilution of the sample, solid urea or Gua-HCl can be added.
Tips:Samples containing urea can be analyzed directly by SDS-PAGE whereas samples containing Gua-HCl must be buffer-exchanged
to a buffer with urea before SDS-PAGE.
Column has clogged:
?Cell debris in the sample may clog the column. Clean the column according to the section Cleaning-in-place.
?It is important to centrifuge and/or filter the sample through a
0.22 μm or a 0.45 μm filter, see Sample preparation. Sample is too viscous:
?If the lysate is very viscous due to a high concentration of host nucleic acid, continue sonication until the viscosity is reduced,
and/or add DNase I to 5 μg/ml, Mg2+ to 1 mM, and incubate on
ice for 10–15 min. Alternatively, draw the lysate through a
syringe needle several times.
Protein is difficult to dissolve or precipitates during purification:
?The following additives may be used: 2% Triton X-100, 2% Tween 20, 2% NP-40, 2% cholate, 1% CHAPS, 1.5 M NaCl,
50% glycerol, 20 mM ?-mercaptoethanol, 1–3 mM DTT or DTE
(up to 5 mM is possible but depends on the sample and the
sample volume), 5 mM TCEP, 10 mM reduced glutathione, 8 M
urea, or 6 M Gua-HCl. Mix gently for 30 min to aid solubilization
of the tagged protein (inclusion bodies may require longer
mixing). Note that Triton X-100 and NP-40 (but not Tween)
have a high absorbance at 280 nm. Furthermore, detergents
cannot be easily removed by buffer exchange.
No histidine-tagged protein in the purified fractions:?Elution conditions are too mild (histidine-tagged protein still bound: Elute with an increasing imidazole gradient or
decreasing pH to determine the optimal elution conditions.
?Protein has precipitated in the column: For the next
experiment, decrease amount of sample, or decrease protein
concentration by eluting with linear imidazole gradient instead
of imidazole steps. Try detergents or changed NaCl
concentration, or elute under denaturing (unfolding)
conditions (add 4–8 M urea or 4–6 M Gua-HCl).
?Nonspecific hydrophobic or other interaction: Add a nonionic detergent to the elution buffer (e.g. 0.2% Triton X-100)
or increase the NaCl concentration.
?Concentration of imidazole in the sample and/or binding buffer is too high: The protein is found in the flow-through
material. Decrease the imidazole concentration.
?Histidine-tag may be insufficiently exposed: The protein is found in the flowthrough material; perform purification of
unfolded protein in urea or Gua-HCl as for inclusion bodies.
?Buffer/sample composition is incorrect: The protein is found in the flowthrough material. Check pH and composition of
sample and binding buffer. Ensure that chelating or strong
reducing agents are not present in the sample at too high
concentration, and that the concentration of imidazole is not
too high.
SDS-PAGE of samples collected during the preparation of the
bacterial lysate may indicate that most of histidine-tagged protein is located in the centrifugation pellet. Possible causes and
solutions are:
?Sonication may be insufficient: Cell disruption may be checked by microscopic examination or monitored by
measuring the release of nucleic acids at A260. Addition of
lysozyme (up to 0.1 volume of a 10 mg/ml lysozyme solution in
25 mM Tris-HCl, pH 8.0) prior to sonication may improve
results. Avoid frothing and overheating as this may denature
the target protein. Over-sonication can also lead to
copurification of host proteins with the target protein.
?The protein may be insoluble (inclusion bodies): The protein can usually be solubilized (and unfolded) from inclusion bodies
using common denaturants such as 4–6 M Gua-HCl, 4–8 M
urea, or strong detergents. Prepare buffers containing 20 mM
sodium phosphate, 8 M urea, or 6 M Gua-HCl, and suitable
imidazole concentrations, pH 7.4–7.6. Buffers with urea should
also include 500 mM NaCl. Use these buffers for sample
preparation, as binding buffer and as elution buffer. For
sample preparation and binding buffer, use 10–20 mM
imidazole or the concentration selected during optimization
trials (including urea or Gua-HCl).
The protein is collected but is not pure (multiple bands on SDS polyacrylamide gel):
?Partial degradation of tagged protein by proteases: Add protease inhibitors (use EDTA with caution, see Table2).
?Contaminants have high affinity for nickel ions: Elute with a stepwise or linear imidazole gradient to determine optimal
imidazole concentrations to use for binding and for wash; add
imidazole to the sample in the same concentration as the
binding buffer. Wash before elution with binding buffer
containing as high concentration of imidazole as possible,
without causing elution of the tagged protein. A shallow
imidazole gradient (20 column volumes or more), may
separate proteins with similar binding strengths. If optimized
conditions do not remove contaminants, further purification
by ion exchange chromotography (HiTrap Q HP or
HiTrap SP HP) and/or gel filtration (Superdex? Peptide,
Superdex 75 or Superdex 200) may be necessary.
?Contaminants are associated with tagged proteins: Add detergent and/or reducing agents before sonicating cells.
Increase detergent levels (e.g. up to 2% Triton X-100 or
2% Tween 20), or add glycerol (up to 50%) to the wash buffer
to disrupt nonspecific interactions.
Histidine-tagged protein is eluted during sample loading/wash:
?Buffer/sample composition is incorrect: Check pH and composition of sample and binding buffer. Ensure that
chelating or strong reducing agents are not present in the
sample at a too high concentration, and that the
concentration of imidazole is not too high.
?Histidine-tag is partially obstructed: Purify under denaturing conditions (use 4–8 M urea or 4–6 M Gua-HCl).
?Column capacity is exceeded: Join two or three HisTrap HP
1 ml columns together or change to a HisTrap HP 5 ml column. 10Intended use
The HisTrap HP is intended for research use only, and shall not be used in any clinical or in vitro procedures for diagnostic purposes.
11Ordering Information
Product No. supplied Code No.
HisTrap HP 5 × 1 ml 17-5247-01
100 × 1 ml1 17-5247-05
1 × 5 ml 17-5248-01
5 × 5 ml 17-5248-02
100 × 5 ml1 17-5248-05 Related products No. supplied Code No.
100 ml 17-5268-02 HiTrap Desalting 5 × 5 ml 17-1408-01
100 × 5 ml1 11-0003-29 PD-10 Desalting Column 30 17-0851-01 HiPrep 26/10 Desalting 1 × 53 ml 17-5087-01
4 × 53 ml 17-5087-02
HisTrap FF 5 × 1 ml 17-5319-01
100 × 1 ml1 17-5319-02
5 × 5 ml 17-5255-01
100 × 5 ml1 17-5255-02 HisTrap FF crude 5 × 1 ml 11-0004-58
100 × 1 ml1 11-0004-59
5 × 5 ml 17-5286-01
100 × 5 ml1 17-5286-02 HisTrap FF crude Kit 1 kit 28-4014-77 HisPrep? FF 16/10 1 × 20 ml 28-9365-51
1Pack size available by special order.
1 One connector included in each HiTrap package.
2 Two, five, or seven stop plugs female included in HiTrap packages depending on the product.
3
One fingertight stop plug is connected to the top of each HiTrap column at delivery.
Accessories
No. Supplied Code No.
1
Tubing connector flangeless/M6 female 1
2 18-1003-68
Tubing connector flangeless/M6 male 1 2 18-1017-98
Union 1/16” female/M6 male 1 6 18-1112-57
Union M6 female /1/16” male 1 5 18-3858-01Union luerlock female/M6 female 2
18-1027-12
HiTrap/HiPrep, 1/16” male connector for ?KTA design
8 28-4010-81Stop plug female, 1/16”2 5 11-0004-64Fingertight stop plug, 1/16”3
5 11-0003-55
Related literature
Code No.Recombinant Protein Purification Handbook, Principles and Methods
18-1142-75Affinity Chromatography Handbook, Principles and Methods
18-1022-29Affinity Chromatography, Columns and Media Selection Guide
18-1121-86Ni Sepharose and IMAC Sepharose, Selection Guide 28-4070-92HiTrap Column Guide
18-1129-81
For local office contact information, visit: https://www.wendangku.net/doc/313264905.html,/contact
GE Healthcare Bio-Sciences AB
Bj?rkgatan 30
751 84 Uppsala
Sweden
https://www.wendangku.net/doc/313264905.html,/hitrap
https://www.wendangku.net/doc/313264905.html,/protein-purification GE Healthcare Europe GmbH Munzinger Strasse 5,
D-79111 Freiburg,
Germany
GE Healthcare UK Ltd Amersham Place
Little Chalfont Buckinghamshire, HP7 9NA
UK
GE Healthcare Bio-Sciences Corp 800 Centennial Avenue
P.O. Box 1327
Piscataway, NJ 08855-1327 USA
GE Healthcare Bio-Sciences KK Sanken Bldg.
3-25-1, Hyakunincho Shinjuku-ku, Tokyo 169-0073 Japan
GE, imagination at work and GE monogram are trademarks of General Electric Company.
?KTA, Drop Design, HiPrep, HisPrep, HisTrap, HiTrap, MidiTrap, MiniTrap, Sephadex, Sepharose, Superdex, and Tricorn are trademarks of GE Healthcare companies.
IMAC Sepharose products, Ni Sepharose products and Fe Sepharose products. These products are covered by US patent number 6 623 655 and equivalent patents and patent applications in other countries.
IMAC Sepharose products and Ni Sepharose products (Histidine-tagged protein purification, Lab products). Purification and preparation of fusion proteins and affinity peptides comprising at least two adjacent histidine residues may require a license under US patent numbers 5,284,933 and 5,310,663, and equivalent patents and patent applications in other countries (assignee: Hoffman La Roche, Inc).
The Tricorn column and components are protected by US design patents USD500856,
USD506261, USD500555, USD495060 and their equivalents in other countries.
All third party trademarks are the property of their respective owners.
? 2006-2009 General Electric Company – All rights reserved.
First published Jan. 2006.
All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. A copy of these terms and conditions is available on request. Contact your local GE Healthcare representative for the most current information.
71-5027-68 AH 03/2009
imagination at work