For technical support, call your local Bio-Rad office or, in the U.S.,
call 1-800-4BIORAD (1-800-424-6723)
Profinia™Native IMAC
and GST Starter Kits
Instruction Manual
(Step-by-Step Guide to
Profinia Operation)
Table of Contents
...
..
Section 1 Introduction............................................................1
1.1 Background ....................................................................1
1.2 Product Information..........................................................1
1.3 Storage Conditions ..........................................................2
Section 2 Native IMAC and GST Starter Kits ........................2
2.1 Native IMAC Component Descriptions ............................2
2.2 GST Component Descriptions..........................................3
Section 3 Equipment and Materials ......................................5
3.1 Profinia Instrument and Accessories ................................5
Section 4 Setup and Running of the Profinia
IMAC Starter Kit ....................................................5
4.1 Reagent Preparation........................................................5
4.2 Instrument Startup, User Interface, and
Method Selection ............................................................6
Section 5 Setup and Running of the Profinia
GST Starter Kit ....................................................10
5.1 Reagent Preparation......................................................10
5.2 Instrument Startup, User Interface, and
Method Selection ..........................................................11
Section 6 Software and Analysis ........................................12
6.1 Viewing Starter Kit Data ................................................12
6.2 Analysis of Purified Protein ............................................13
Section 7 Shutdown, Cleaning, Storage Sequence,
and Data Export ..................................................14
Section 8 Frequently Asked Questions and
Troubleshooting ..................................................15
Section 9 Legal Notices ......................................................16
Section 10 Ordering Information ..........................................17
Section 1
Introduction
1.1 Background
The Profinia protein purification system includes a complete line of
consumables to use with an automated, user-friendly instrument for the
purification of immobilized metal affinity chromatography (IMAC) and
glutathione-S-transferase (GST) affinity-tagged proteins. The system has
been designed to use standard purification buffers, resins, and reagents
packaged into an easy-to-use kit. The two most common purification
applications, native IMAC and GST, have been packaged into starter kits
that contain all components necessary to purify a control affinity-tagged
protein. The buffers are supplied as concentrates and are diluted by the
instrument; no predilution is required. Profinia IMAC and GST starter kits
can be used after instrument installation and setup to familiarize the user
with the instrument or when troubleshooting purification problems. See
Section 10 for starter kit information and the complete list of Profinia products.
1.2 Product Information
Profinia starter kits are used to get started with the Profinia protein
purification system. The kits also help diagnose proper operation of the system. The Profinia starter kits contain a lyophilized 51 kD dual-tagged protein
that contains an N-terminal GST sequence tag and a C-terminal tag with
histidine (His) residues.* Once resuspended in diluted equilibration buffer,
the lysate yields sufficient protein for two independent purifications. Each
kit contains a complete set of buffers and solutions (affinity, desalting,
cleaning, and storage solutions), reagents (glutathione), and cartridges (1 ml
affinity paired with a 10 ml desalting) required for two affinity purifications
only. However, additional Bio-Scale™Mini IMAC or GST cartridges can be
purchased separately, as the buffers provided in the starter kits are
sufficient to perform 10 purification runs. See Proper Storage of Solutions
and Kits on page 15. Table 1 summarizes the reagents contained within the
two starter kits available for the Profinia system.
* The C-terminal histidine tag contains alternating histidine units and is not recognized by
6x histidine antibodies.
1
2
Table 1. Profinia IMAC and GST starter kits.
Kit Catalog # Solutions Reagents Cartridges
Purification Desalting Storage
Buffers Buffer Solutions Lysate Glutathione Affinity Desalting
Native IMAC 620-0229
✔✔✔✔ ✔✔
starter kit
GST starter kit 620-0230 ✔✔✔✔ ✔ ✔✔
1.3 Storage Conditions
Profinia starter kits are shipped at room temperature and can be safely
stored for short periods (up to 2 weeks) at room temperature. For long-term
storage, the kits should be stored at 4°C. The individual components and
outside kit box labels provide exact expiration dates.
Section 2
Native IMAC and GST Starter Kits
2.1 Native IMAC Component Descriptions
The affinity buffers provided in the native IMAC kit are formulated from
potassium salts and buffers, and contain increasing concentrations of
imidazole for optimized washing and eluting of His-tagged proteins. Native
IMAC buffers are used for proteins soluble in
E. coli
lysates and that have
accessible tags containing histidine residues. Table 2 provides a list of
buffer compositions. The native lysis buffer is used for preparing
sample lysates; is not used in the instrument. This buffer can be used for
subsequent purifications of target protein outside of the starter kit
applications.
The IMAC cartridges are shipped in 20% ethanol and are packed with
1 ml Profinity™IMAC resin. The desalting cartridges are provided in
20 mM Bis-Tris, pH 6.5 with 0.05% azide as a preservative. The control
lysate is provided as a lyophilized sample prepared from 50 ml
E. coli
culture. When resuspended in 12 ml of 1x IMAC lysis buffer, ~4 mg of
tagged target protein can be purified from a 5 ml sample load. All
components supplied with the native IMAC starter kit are shown in Table 2.
Table 2. Formulations for buffers and solutions provided in the
native IMAC starter kit.
Profinia Solution Supplied Concentrated Formulation Working 1x Formulation Volume Position #
As
Native IMAC 2x 600 mM KCl, 100 mM KH
2PO4
, 300 mM KCl, 50 mM KH2PO4,
lysis buffer 10 mM imidazole, pH 8.0 5 mM imidazole, pH 8.0 125 ml N/A
Native IMAC 2x 600 mM KCl, 100 mM KH
2PO4
, 300 mM KCl, 50 mM KH2PO4,
wash buffer 1 10 mM imidazole, pH 8.0 5 mM imidazole, pH 8.0 125 ml 1
Native IMAC 2x 600 mM KCl, 100 mM KH
2PO4
, 300 mM KCl, 50 mM KH2PO4,
wash buffer 2 20 mM imidazole, pH 8.0 10 mM imidazole, pH 8.0 100 ml 2
Native IMAC 2x 600 mM KCl, 100 mM KH
2PO4
, 300 mM KCl, 50 mM KH2PO4,
elution buffer 500 mM imidazole, pH 8.0 250 mM imidazole, pH 8.0 100 ml 3
Desalting buffer 5x 685 mM NaCl, 13.5 mM KCl, 137 mM NaCl, 2.7 mM KCl,
21.5 mM Na
2
HP04, 4.3 mM Na2HP04,
40.5 mM KH
2
HP04, 8.1 mM KH2P04, pH 7.4 200 ml 4
pH 7.0 (pH 7.4 upon dilution)
Cleaning solution 1 2x 1,000 mM NaCl, 500 mM NaCl,
100 mM Tris, pH 8.0 50 mM Tris, pH 8.0 125 ml 5
Cleaning solution 2 4x 2,000 mM NaCl, 500 mM NaCl,
400 mM NaOAc, pH 4.5 100 mM NaOAc, pH 4.5 125 ml 6
Storage solution 2x 4% C
6H5CH2
OH 2% C6H5CH2OH
(benzyl alcohol) (benzyl alcohol) 200 ml 7
2.2 GST Component Descriptions
The lysis and wash buffers contained in the GST kit are formulated
from sodium salts, phosphate buffers, and EDTA, and provide optimized
binding, washing, and eluting of GST-tagged proteins. EDTA is included
as a chelating compound that protects against metalloproteases, which can
be present in
E. coli
lysates. GST fusion proteins must be in a native
conformation prior to purification. The buffers used in this system are
designed for the purification of proteins that are soluble in
E. coli
lysates
and that have accessible, native GST sequence tags. Table 3
provides a list of buffer compositions. The lysis buffer is used for preparing
sample preparations; it is not used in any of the instrument ports. This
buffer can be used for subsequent purifications of target protein outside of
the starter kit applications.
The GST cartridges are shipped in 20% ethanol and are packed with
1 ml Profinity GST resin. The desalting cartridges are provided in 20 mM
Bis-Tris, pH 6.5 with 0.05% azide as a preservative. The vial of lyophilized
lysate is described in sections 1.2 and 2.1, and when resuspended in 12 ml
1x GST lysis buffer, provides adequate sample for two independent
purification runs.
3
Table 3. Formulations for buffers and solutions provided in the GST
kits.
Profinia Supplied
Solution AsConcentrated Formulation Working 1x Formulation Volume Position #
GST 2x 300 mM NaCl, 20 mM 150 mM NaCl, 10 mM Na
2
HPO4,
lysis buffer Na
2
HPO4, 10 mM EDTA, 5 mM EDTA, pH 7.4 125 ml N/A
pH 7.4
GST 2x 300 mM NaCl, 20 mM 150 mM NaCl, 10 mM Na
2
HPO4,
wash buffer Na
2
HPO4, 10 mM EDTA, 5 mM EDTA, pH 7.4 200 ml 1
pH 8.0
GST 2x 40 mM glutathione, 200 mM Tris, 20 mM glutathione, 100 mM Tris,
elution buffer 10 mM EDTA, pH 8.0 5 mM EDTA, pH 8.0 100 ml 3
Desalting 5x 685 mM NaCl, 13.5 mM KCI, 137 mM NaCl, 2.7 mM KCl,
buffer 21.5 mM Na2HPO4, 40.5 mM 4.3 mM Na2HPO4, 8.1 mM
KH
2
HPO4, pH 7.0 KH2PO4, pH 7.4 200 ml 4
(pH 7.4 upon dilution)
Cleaning 2x 1,000 mM NaCl, 500 mM NaCl, 50 mM Tris,
solution 1 100 mM Tris, pH 8.0 pH 8.0 125 ml 5
Cleaning 4x 2,000 mM NaCl, 500 mM NaCl,
solution 2 400 mM NaOAc, pH 4.5 100 mM NaOAc, pH 4.5 125 ml 6
Storage 2x 4% C
6H5CH2
OH 2% C6H5CH2OH
solution (benzyl alcohol) (benzyl alcohol) 200 ml 7
4
Section 3
Equipment and Materials
3.1 Profinia Instrument and Accessories
Section 4
Setup and Running of the Profinia IMAC
Starter Kit
4.1 Reagent Preparation
Open the starter kit and remove the vial of lyophilized lysate. Dilute
6 ml of IMAC lysis buffer with 6 ml distilled water to make 12 ml of a 1x
solution. Resuspend the vial of lyophilized lysate with the 12 ml 1x lysis
buffer, swirl to mix, and let stand 5 min at room temperature to
completely dissolve. Note: Do not use lysis buffer from the GST starter kit
for IMAC purification because 10 mm EDTA is included in the GST lysis
buffer and will strip the nickel from the IMAC cartridge. Transfer 6 ml of the
resuspended sample to a 15 ml conical tube and place on ice until ready
to start purification. The remaining 6 ml of resuspended sample can be
frozen for later use and thawed once prior to purification. The frozen
sample should be used within 3 months.
The IMAC buffers and cartridges are ready for insertion into the
instrument and require no advanced preparation.
5
The instrument comes packaged
with nearly all materials necessary
for the setup and purification of the
control protein provided in the starter
kit. The instrument is shipped with all
the key bottles and plasticware
needed to run the system: waste
and diluent bottles, a bottle starter
pack, and a cleaning tray. See
Figure 1 for a picture of the
accessories. Materials not provided
in the package include standard 15
ml or 50 ml tubes that are required
for the sample and collection ports.
Most standard 15 and 50 ml tubes fit
the required dimensions of the
Profinia system.
Fig. 1. Accessories for the Profinia instrument.
Items shown include waste and diluent bottles (top),
bottle starter pack (left), sample cooling device (right),
cleaning tray (bottom), sample loop (bottom left), 15 ml
and 50 ml sample lids and buffer bottle lids (middle).
4.2 Instrument Startup, User Interface, and Method
Selection
Turn on the instrument using the recessed on/off switch at the right
lower panel. The Profinia home screen will appear. Make sure the cleaning
tray is in place and that filtered water is in buffer positions 1–7. Choose
Data/Utilities, Diag/Maint Functions, and Clean All Inlet and Outlet Lines.
This will flush ethanol or air from the system prior to starting a
chromatography run. After the system is rinsed press Home to return to
the Profinia home screen. Choose Bio-Rad Methods. The user interface
guides users through the setup and running of the preprogrammed
method. A brief description of each key screen and necessary actions to
take to perform a purification using the 51-D lysate are outlined below.
Select Method System Screen
The Select Method System screen
allows the user to choose among
different purification systems (IMAC,
GST, Desalting, Protein A&G, eXact).
(Figure 2)
•IMAC
Select Method Type and Options
Screen
The Select Method Type and Options
screen allows the user to quickly
choose the method of purification,
number of samples to be purified,
and volumes for the affinity and/or
desalting cartridges. The selection in
Figure 3 are.
• Method type: Native IMAC +
Desalting
• Sample type: 1 sample
• Cartridge selection: 1 ml + 10 ml
6
Fig. 3. Method selection for native IMAC
and desalting application.
Fig. 2. Method selection for affinity
purification.
7
Select Sample Parameters Screen
The Select Sample Parameters screen
gives users the option of increasing/
decreasing the rate of flow or the
number of column volumes of wash
used. The selections in Figure 4 are.
• Flow rate: Standard*
• Wash time: Standard
* Standard flow rates for IMAC purifications are
2 ml/min. Proteins that are very large (>100 kD),
have His residues that are not fully exposed, or require
cold room purifications, typically bind with slower
kinetics. In these situations, the slow flow rate
option (0.5 ml/min) increases binding efficiency.
Enter Run and Sample Information
Screen
Users can customize the method by
adding user-specific details that
distinguish samples from one another
(Figure 5). Such details may include
user and run name, sample name
and volume, and A
280
of a 1 mg/ml
protein solution. Selections in Figure 5
are:
• Username: Anonymous
• Run name: UNTITLED
• Lot tracking: BLANK
• Sample 1 name: 3/4 vial51kd
• Volume: 5.00 ml
•A
280
of 1 mg/ml*: 1.33
*All proteins absorb UV light based upon amino acid
composition. The A
280
of a 1 mg/ml protein solution
can be calculated if the specific amino acid sequence
is known. For 51-D in the control lysate, the OD
280
of a
1 mg/ml solution is 1.33. By entering this value,
the Profinia instrument automatically calculates and
converts the concentration of the purified protein from
the absorbance values. The concentrations given by
the Profinia system are estimates of the purified protein.
Analytical determination of protein concentration using
a spectrophotometer or Bradford analysis following
purification is always good practice.
Fig. 5. Run and sample information
selection screen.
Fig. 4. Method selection for sample flow
rate and wash times.
Install Reagents, Sample, and
Fraction Tubes Screen
Installation of reagents, samples,
and fraction tubes is specific to the
method type chosen. On one screen,
necessary components are shown if
needed (or crossed out, Figure 6) so
that the user can quickly and easily
proceed with the setup of the
instrument.
Reagent and bottle icons: Install the
buffer bottles, sample tubes, waste
and water diluent containers, and
collection tubes as illustrated by the
icons on the screen. The number of
buffer or water volume indicates the
minimal volume needed for this size
of the containers to avoid introducing
of bubble due to insufficient fluid.
Select Cartridge Status Screens
a user interface screen that describes
cartridge installation shown in
Figure 7.
Select Cartridge Status: The
system must be primed with buffer
before installing cartridges to
prevent air or storage ethanol from
being pumped into the cartridges.
On the Select Cartridge Status
menu choose NO if the lines have
not yet been primed and cartridges
are not installed. Pressing the NO
button brings you to the Prepare
For Cartridge Line Priming screen.
Choose YES only if you have just
completed a run of the same type
and have not removed the
chromatography cartridges.
Choosing YES will send you to the
Start Run Screen.
8
Fig. 6. Reagents, sample, and fraction
tubes screen.
Fig. 7. Cartridge installation screens.
Prepare For Cartridge Line
Priming: This screen directs users to
make sure the fittings for both
cartridge positions 1 and 2 are
connected without any cartridges in
place (Figure 8). Pressing Start
primes the cartridge lines with buffer
at high flow rate and then brings
users to the Install Cartridges screen.
Install Bio-Scale Mini Cartridges:
Disconnect the upper and lower
cartridge holder fittings. Unscrew the
bottom luer cap of a cartridge and
attach it by rotating; clockwise.
Remove the top luer cap of the
cartridge. Attach it by lowering the top
holder and tightening the fitting into
place (Figure 9).
9
Fig. 9. Installation of cartridges on the Profinia
system.
Fig. 8. Instruction screen for cartridge line
primary before cartridge installation.
Run Screens
A run is commenced by pressing the Start button. Once started, the
run in progress screen provides the approximate elution time for the
sample (typically 37 minutes for a demo run). Return to the instrument at
this time to retrieve the purified sample. (Figure 10)
Start run: This screen initiates the
purification run and notifies the user
how to view data in real time with
the Profinia data collection software
package.
Run in progress: A Run in Progress
screen is displayed for each method
selection. Features such as a status
bar, current step in the protocol
(for example, Equilibrate, Wash 1,
Elute), and expected time to sample
elution are all specified, so it is easy
to identify where the sample is at
any given point in the protocol.
Section 5
Setup and Running of the Profinia GST
Starter Kit
5.1 Reagent Preparation
Open the starter kit and remove the vial of lyophilized lysate. Dilute 6 ml
GST lysis buffer 1:1 with 6 ml distilled water to make 12 ml of a 1x solution.
Resuspend the vial of lyophilized lysate with the 12 ml of 1x lysis buffer,
swirl to mix, and let stand 5 min at room temperature to completely dissolve.
Transfer 6 ml of the resuspended sample to a 15 ml
10
Fig. 10. Start run and run in progress screens.
conical tube and place on ice until ready to start the purification. The
remaining 6 ml of resuspended sample can be frozen for later use and
thawed once prior to purification. The frozen sample should be used within
6 months.
Remove the bottle of GST elution buffer and vial of reduced glutathione.
Transfer 10 ml elution buffer to the vial of glutathione, shake to mix, and
let stand 5 min at room temperature to completely dissolve. Transfer the
resuspended glutathione to the 125 ml bottle of elution buffer. The elution
buffer (2x) now contains 40 mM glutathione, which is necessary to elute
GST-tagged proteins. The remaining bottles of GST buffers and cartridges
require no advance preparation.
5.2 Instrument Startup, User Interface, and Method
Selection
Turn on the instrument using the recessed on/off switch at the right
lower panel. Make sure the cleaning tray is in place and that filtered water
is in buffer positions 1–7. Choose Data/Utilities, Diag/Maint Functions, and
Clean All Inlet and Outlet Lines. This will flush ethanol or air from the system
prior to starting the chromatography run. After the system is rinsed press
Home to return to the Profinia home screen. Choose Bio-Rad Methods.
The user interface for the GST starter kit is similar to that described in
Section 12. Choose GST on the select method screen. The method
screen for the GST method type and options should look like Figure 11.
Select Method Type and Options
Screen
The Select Method Type and Options
screen allows the user to quickly
choose the method of purification,
number of samples to be purified, and
the column volume (ml) for the affinity
and/or desalting cartridges. The
selections in Figure 11 are:
• Method type: GST + Desalting
• Sample type: 1 sample
• Cartridge selection: 1 cartridge
11
Fig. 11. Method selection for GST application.
Section 6
Software and Analysis
6.1 Viewing Starter Kit Data
The standard software report presents key sample data collected throughout
the run. The sample name, fraction location and volume, and estimated
total protein eluted into collection tube D are displayed in the table in Figure 13.
The report can be formatted and printed in a variety of formats. The table
in Figure 13 highlights a partial section of the report.
Fig. 13. Standard software report.
12
Software is an optional accessory to
the Profinia instrument and is
required to visualize chromatographic
data from the purification profile. A
representative chromatogram from
the 51-D control lysate purification is
shown in Figure 12.
The y-axis displays the absorbance
units (AU) and the x-axis displays
the run time (h:mm:ss). The first
peak traced by UV-1 illustrates the
unbound protein found in all fractions: load (L), flowthrough (FT),
wash 1 and wash 2 (W-1, W-2), and
elution. The second peak traced by
UV-1 contains the affinity purified
protein, which is then diverted to the
desalting cartridge. The final peak
traced by UV-2 contains the purified
and desalted protein collected in the
elution fraction. A gel showing the
protein from each of the main
purification steps is shown in Figure 13.
Fig. 12. Representative chromatogram
displayed on Profinia software.
6.2 Analysis of Purified Protein
13
The standard report also summarizes
key run parameters such as time to
elution, flow rate, wash time, and
volume loaded. If the software
package is not part of your Profinia
system, the end of run summary
screen presents the key sample data
directly on the user interface screen
(Figure 14).
Fig. 14. Data report screen on user interface
screen.
Purified affinity-tagged proteins are
typically analyzed by gel
electrophoresis. For IMAC
purifications, samples of starting
material (L, resuspended lysate, or
load), flowthrough (FT, tube A),
wash 1 (W-1, tube B), wash 2 (W-2,
tube C), and purified protein (E, tube
D) are diluted 1:7 in Laemmli buffer,
boiled, and 10–20 µl is then loaded
onto a gel. The IMAC buffers contain
potassium salts and must be diluted
at least 1:7 to prevent potassium-SDS
precipitates from forming in Laemmli
buffer. The gel in Figure15 shows a
representative purification from the
51-D IMAC purification.
Fig. 15. Gel analysis of purified 51-D with
native IMAC.
LFTW-1W-2E
Section 7
Shutdown, Cleaning, Storage Sequence,
and Data Export
After the run is completed, the instrument will instruct the user to begin
the proper cleaning and storage sequence. To maintain the instrument
in proper working order and to prevent mechanical breakdown,
always follow the short- or long-term storage commands. Short-term
storage is used when the instrument will be run within 2 days, and
long-term storage is appropriate when the instrument will be used in 2 or
more days.
Prepare System to Clean Lines
• Remove sample tubes, install tubes filled with water, and install the
cleaning tray. Press Start to begin the cleaning process. Refer to
Figure 16 for the screen associated with this command
Fig. 16. Preparing the system for cleaning.
Export Data
• Once the sample and fraction lines are cleaned, the data can be
exported through a USB drive. Install a USB drive into the USB port
located on the front of the instrument, and press Export Data
• The exported data can be viewed on any PC using the Profinia
software package
Short-Term/Long-Term Wash
• Choose Short-Term Wash to store for <2 days. Choose Long-Term
Wash to store for >2 days
14
• Remove the buffer bottles in positions 1–7 and replace with bottles
that contain water (the bottles in the starter pack should be used for
this purpose)
• Remove the cartridges and connect the upper and lower cartridge
fittings. Press Start to begin the cleaning cycle
• If the long-term wash cycle is chosen, install 20% ETOH in buffer
positions 1–8, sample positions 1 and 2, and at the DI water inlet. The
ethanol will be used as a long-term storage solution in the lines
Section 8
Frequently Asked Questions and
Troubleshooting
For an extensive and comprehensive list of troubleshooting tips for
protein expression, sample prep, purification, and analysis, refer to the
Profinia Protein Purification Instrument Instruction Manual. Questions and
tips below focus on the use of the starter kits and solutions.
Proper Storage of Solutions and Kits
• Both kits can be stored at 22°C (room temperature) for short periods
of time, either upon receipt or during normal usage. Due to the sensi-
tivity of some of the more labile reagents (control lysate,
glutathione, GST cartridges), the kits should be kept at 4°C for
long-term storage
• Once opened and used with the instruments, all reagents should be
stored at 4°C for up to 3 months. After insertion into the instrument,
the solutions are no longer sterile and require 4°C storage
• After opening and use with the instrument, check the solutions for
particulates and clarity before reusing. If there is any indication of
particulates, contamination, or microbial growth, the solutions should
be discarded
Use of Excess Reagents
• The limiting reagent in the kit is the control lysate, which is sufficient
for two purifications yielding ~4 mg of target protein. The lysate can be
loaded at lower or higher amounts to mimic the purification of low-
level or high-level target proteins
15
• There are sufficient buffers and reagents provided for ten 1 ml
purification runs. If it is desirable to use the remaining reagents on
proteins other than the supplied control, the reagents should be stored
at 4°C and used within 6 months. The elution buffer, which contains
glutathione, must be stored at –20°C and used within 6 months. The
solution should be thawed only once
Reuse of Cartridges
• The 1 ml affinity and 10 ml desalting cartridges provided in the starter
kit can be used up to five times. Since the lysate provided in the kit is
sufficient for only two purifications, cartridges could theoretically be
used three more times. However, as with any type of affinity
purification, it is suggested that individual cartridges be dedicated to
the purification of a unique protein, in this case the control lysate
protein. Cross-contamination can always pose a risk if more than one
protein is purified over the same cartridge
• If the user chooses to use the same 1 ml affinity and 10 ml desalting
cartridge for a different protein, Section 7 provides the appropriate
cleaning and storage protocols for cartridge reuse
• Alternatively, and if cross-contamination is a concern, Bio-Scale™
Mini IMAC or GST cartridges may be purchased separately can
buffers provided in the starter kit are provided in sufficient quantity to
perform 10 purification runs
Section 9
Legal Notices
Purification and preparation of fusion proteins and affinity peptides
containing at least two adjacent histidine residues may require a license
under U.S. patent 5,284,933 and U.S. patent 5,310,663, including foreign
patents (assignee Hoffman-LaRoche).
Expression and purification of GST fusion proteins may require a
license under U.S. patent 5,654,176 (assignee Chemicon International).
16
Section 10
Ordering Information
Catalog # Description
Profinia Systems
620-1005 Profinia Instrument with Accessory Kit and Native IMAC
Starter Kit, 100–240 V
620-1006 Profinia Instrument with Accessory Kit and GST Starter Kit,
100–240 V
620-1010 Profinia System with Software, Accessory Kit and Native
IMAC Starter Kit, 100–240 V
620-1011 Profinia System with Software Accessory Kit and GST
Starter Kit, 100–240 V
Profinia Accessories
620-0010 Profinia Software with usb cable
620-0401 Profinia Cooling Accessory
620-0402 Profinia Desalting Sample Loop, 2.0 ml
620-0403 Profinia Desalting Sample Loop, 10.0 ml
620-0404 Profinia Instrument Inline Filter Assembly Pack
620-0410 Profinia Instrument Accessory Kit, includes cleaning tray, inline
filter assembly replacement, 8 buffer lids, 2 x 50 ml sample lids, 2 x
15 ml lids, bottle starter pack, waste/diluent bottle set
620-0231 Bottle Starter Pack, 4 x 125 ml and 4 x 250 ml bottles and 8
buffer lids
620-0411 Profinia pH Monitor Kit with mounting accessory kit, includes
pH electrode, flow cell, mounting accessories
732-0112 Adaptor Kit, connecting to GE Healthcare, Pierce pre-packed
cartridges
Profinia Purification Kits
620-0225 Profinia Native IMAC Purification Kit, 1 ml, includes Profinia
native IMAC buffer kit, 2 x 1 ml IMAC and 2 x 10 ml desalting
cartridges
620-0235 Profinia Native IMAC Purification Kit, 5 ml, includes 2 Profinia
native IMAC buffer kits, 1 x 5 ml IMAC and 1 x 50 ml desalting
cartridges
620-0226 Profinia GST Purification Kit, 1 ml, includes Profinia GST buffer
kit, 2 x 1 ml GST and 2 x 10 ml desalting cartridges
620-0236 Profinia GST Purification Kit, 5 ml, includes 2 Profinia GST
buffer kits, 1 x 5 ml GST and 1 x 50 ml desalting cartridges
17
Catalog # Description
620-0228 Profinia Desalting Purification Kit, 10 ml, includes desalting and
cartridge cleaning buffer kit, 2 x 10 ml desalting cartridges
620-0238 Profinia Desalting Purification Kit, 50 ml, includes 2 desalting
and cartridge cleaning buffer kits, 1 x 50 ml desalting cartridge
Profinia Buffer Kits
620-0221 Profinia Native IMAC Buffer Kit, includes purification buffers,
cleaning and storage solutions; sufficient for 10 applications for
1 ml cartridge
620-0223 Profinia GST Buffer Kit, includes purification buffers, cleaning
and storage solutions, glutathione reagent; sufficient for 10
applications for 1 ml cartridge
620-0224 Desalting and Cartridge Cleaning Buffer Kit, includes desalting
buffer, cleaning and storage solutions; sufficient for 10 applications
for 10 ml cartridge
Profinia Starter Kits
620-0229 Profinia IMAC Starter Kit, includes Profinia native IMAC buffer kit,
1 x 1 ml IMAC and 1 x 10 ml desalting cartridges, E. coli lysate
620-0230 Profinia GST Starter Kit, includes Profinia GST buffer kit, 1 x 1 ml
GST and 1 x 10 ml desalting cartridges, E. coli lysate, glutathione
reagent
Profinia Reagents
620-0203 His Antibody, 100 µl, 1 mg/ml
620-0204 GST Antibody, 100 µl, 1 mg/ml
620-0233 His and GST Purification
E. coli
Control Lysate, Lyophilized
620-0202 Glutathione Pack, 1.23 g
Bio-Scale™ Mini Affinity and Desalting Cartridges
732-4610 Bio-Scale Mini Profinity IMAC Cartridges, 5 x 1 ml
732-4612 Bio-Scale Mini Profinity IMAC Cartridge, 1 x 5 ml
732-4614 Bio-Scale Mini Profinity IMAC Cartridges, 5 x 5 ml
732-4620 Bio-Scale Mini Profinity GST Cartridges, 5 x 1 ml
732-4622 Bio-Scale Mini Profinity GST Cartridge, 1 x 5 ml
732-4624 Bio-Scale Mini Profinity GST Cartridges, 5 x 5 ml
732-5304 Bio-Scale Mini Bio-Gel
®
P-6 Desalting Cartridges, 5 x 10 ml
732-5312 Bio-Scale Mini Bio-Gel P-6 Desalting Cartridge, 1 x 50 ml
732-5314 Bio-Scale Mini Bio-Gel P-6 Desalting Cartridges, 5 x 50 ml
732-4600 Bio-Scale Mini Aff-Prep Protein A Cartridges, 5 x 1 ml
18
Catalog # Description
732-4602 Bio-Scale Mini Affi-Prep Protein A Cartridge, 1 x 5 ml
732-4200 Bio-Scale Mini UNOsphere SUPrA™ Cartridge, 1 x 1 ml
732-4201 Bio-Scale Mini UNOsphere SUPrA Cartridges, 5 x 1 ml
732-4202 Bio-Scale Mini UNOsphere SUPrA Cartridge, 1 x 5 ml
732-4646 Bio-Scale Mini Profinity eXact Cartridges, 2 x 1 ml
732-4647 Bio-Scale Mini Profinity eXact Cartridges, 4 x 1 ml
732-4648 Bio-Scale Mini Profinity eXact Cartridge, 1 x 5 ml
19
Bio-Rad
10006057 Rev E
Laboratories, Inc.
Life Science
Group
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Web site www.bio-rad.com USA 800 4BIORAD
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Finland 09 804 22 00
Greece 30 210 777 4396 Hong Kong 852 2789 3300
Hungary 36 1 455 8800 India 91 124 4029300 Israel 03 963 6050
Italy 39 02 216091 Japan 03 6361 7000 Korea 82 2 3473 4460
Mexico 52 555 488 7670 The Netherlands 0318 540666
New Zealand 0508 805 500 Norway 23 38 41 30 Poland 48 22 331 99 99
Portugal 351 21 472 7700 Russia 7 495 721 14 04
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