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QuickPick solutions for protein fractionation/purification
Each of Bio-Nobiles QuickPick protein kits contain reagents and validated
protocols for a particular method.
Methods are currently available for CM and DEAE weak ion exchange,
immobilized metal affinity chromatography (IMAC),
and glutathione-S-transferase (GST) fusion protein purification.
The kits are optimized for use with the PickPen® 1-M and 8-M magnetic tools as
well as for MagRo robotic workstations with fast and easy operation procedures.
Benefits of Bio-Nobile protein purification solutions
Fast
- Rapid 5-12 minutes
protocols ensure high-quality product
- Short hands-on time
- Move particles, not liquids, and avoid time-consuming aspirating and
dispensing steps
Reliable
- Robust protocols
- Reproducible results
- High-quality purified product
Scalable and flexible
- Easy solution for various sample volumes
- Use with PickPen magnetic tools or MagRo workstation
Wide range of QuickPick product formats
QuickPick Ready-to-use kits
QuickPick protein fractionation and purification ready-to-use kits are
optimized for constant sample volumes to be used with the PickPen 1-M and
PickPen 8-M magnetic tools. These ready-to-use kits contain reagents and
consumables needed for the purification. All of these kits are available in 8
and 48 preparations kit sizes except IMAC Plus which is available in 8
preparation kit size.
QuickPick Scalable kits (SML)
QuickPick IMAC scalable kit can be used for variable sample sizes. The sample
volumes may be scaled up and down, giving the yield of high-quality protein
appropriate for the intended downstream application.
QuickPick Bulk reagents (XL)
QuickPick IMAC reagents are also available for custom and bulk configurations.
Bio-Nobile ion exchange methods (CM and DEAE)
The weak ion exchange methods in
Bio-Nobile QuickPick kits are based on magnetizable
cellulose particles coated with carboxymethyl (CM) or diethyl aminoethyl (DEAE)
groups. Different types of proteins will bind to the CM or DEAE coated magnetic
particles with affinities that depend on both the conditions used and the types
and number of individual charged groups. In typical protein purification using
ion exchange chromatography a mixture of many proteins is used as a sample.
After unbound molecules are washed away conditions can be stepwise adjusted,
such as by altering the pH and salt concentration to release the molecule(s) of
interest from the particles.
High quality purifications using ion exchange methods
Not only do PickPen methods allow high quality purifications, they also allow
flexibility in working methods. You can use the PickPen magnetic tool to
concentrate a protein. This is illustrated below for
the protein, cytochrome c.
| Sample containing cytochrome c |
Cytochrome c removed |
Cytochrome c resuspended |
 |
| 150 μl |
150 μl |
25 μl |
Cytochrome c in I mg/ml solution captured on magnetic particles using the
QuickPick CM kit and resuspended in a smaller
volume.
| Kit |
CM |
DEAE |
 |
Sample materials
- bacterial cells
- yeast cells
- cultured cells
- animal tissue
- plant tissue
|
Cells from 5 ml cultivation lysed in 500 µl
wash buffer and diluted 1:5
500 mg / 1 ml wash buffer
106 cells / 0.5 ml wash
buffer
250 mg / 1 ml wash buffer
250 mg / 1 ml wash buffer |
Cells from 5 ml cultivation lysed in 500 µl wash buffer and diluted 1:5
500
mg / 1 ml wash buffer
106 cells / 0.5 ml wash buffer
250 mg / 1 ml wash
buffer
250 mg / 1 ml wash buffer |
| Sample size |
300-1000 µl |
300-1000 µl |
Processing time for
- one sample
- four
samples
|
<10 min
30-40 min
|
<10 min
30-40 min
|
| Elution volume per reaction |
20-100
µl |
20-100
µl |
|
Yield per prep |
60 ± 6 µg for aprotinin |
60 ± 6 µg for BSA |
Download QuickPick CM or DEAE protocol
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Bio-Nobile immobilized metal ion affinity chromatography (IMAC)
QuickPick IMAC magnetic particles are formed from magnetizable agarose particles
which affords an extensive porous surface area especially preferred in the
purification of histidine tagged proteins from cell lysates. The transfer of
magnetic particles with the PickPen magnetic tool enables rapid purification of histidine tagged proteins with suitable binding and elution characteristics. The
methodology used abolishes the need for sample pre-treatment or use of
time-consuming column technologies.
High quality purifications using QuickPick IMAC
The performance of the QuickPick IMAC kit in purifying His-tagged
protein specifically is illustrated by the example below in which
E.coli His-tagged glutathione-S-transferase (HT-GST) was isolated
using the QuickPick IMAC kit.
The cells were lysed in 20 mM Tris-HCl buffer, pH 7.0 before
purification.
M = Molecular weight marker
1 = Cell lysate
2 = Lysate after MP treatment
3 = 5 µl QuickPick IMAC eluate
4 = 7.5 µl QuickPick IMAC eluate
5 = 10 µl QuickPick IMAC eluate
Purity checked by analyzing 5, 7.5 and 10 µl of eluate on SDS-PAGE gel. The HT-GST protein band is clearly seen for each
of the three eluate lines.
Higher yields by using the QuickPick IMAC Plus kit
The QuickPick IMAC Plus kit is designed for applications which demand higher yields
of protein. As shown below, a yield of about 0.5 mg (~30 kDa protein) is obtainable
from a single purification.
Yield obtained using QuickPick IMAC Plus kit
| Kit |
IMAC |
IMAC Plus |
|
Sample materials
|
Bacterial cells:
E. coli recombinant protein expression:
Cells from 5 ml cultivation lysed in 500 µl wash buffer |
Bacterial cells:
E. coli recombinant protein expression:
Cells from 10-40 ml cultivation lysed in 3 ml wash buffer |
| Sample size |
300-1000 µl |
2 -4 ml |
Processing time for
- one sample
- four
samples
|
<10 min
30-40 min
|
≥10 min (depends on binding time used)
|
| Elution volume per reaction |
20-100
µl |
300-600
µl |
|
Yield per prep |
30 ± 3 µg for HT-GST |
Up to 0.5 mg for HT-GST |
Download QuickPick IMAC protocol
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Bio-Nobile QuickPick GST
kit
QuickPick GST Magnetic Particles are formed from magnetic agarose particles
which provide an extensive porous surface area especially suitable for the
purification of recombinant GST-fusion proteins from cell lysates. The transfer
of magnetic particles with PickPen technology enables rapid purification of
recombinant GST-fusion proteins. The methodology removes the need of using
time-consuming column technology.
High quality purifications using QuickPick GST kit
The performance of the QuickPick GST kit in purifying a GST-fusion protein is
illustrated by the example below in which GST-Btk SH3 (a GST fusion protein of
Brutons tyrosine kinase SH3 domain) expressed in E. coli was isolated using the QuickPick GST kit. The cells were lysed in 20 mM Tris-HCl buffer, pH 7.0 before
purification.
1 = LMW
marker
2 = E. coli cell lysate
3-6 = 3μl eluate from 4
parallel samples
Purified GST-Btk SH3 protein on SDS-PAGE gel
| Kit |
GST |
|
Sample materials
|
Bacterial cells:
E. coli
recombinant protein expression:
Cells from
3-10 ml cultivation lysed in 800 µl wash buffer. |
| Sample size |
800 µl |
Processing time for
- one sample
- four
samples
|
<10 min
30-40 min
|
| Elution volume per reaction |
70-100
µl |
|
Typical yield |
Up to 40
µg/prep (GST) |
| Purity |
> 90% |
Download QuickPick GST for
PickPen® 1-M manual tool
protocol
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