• ABOUT US
  • OUR SERVICES
  • SERVICE FEES
  • PROTOCOLS
  • INSTRUMENTS
  • CONTACT US

© Copyright 2011 rPPC at CLP of Northwestern University.

ABOUT US

Welcome to the Recombinant Protein Production Core!

The Recombinant Protein Production Core (rPPC) provides quality controlled recombinant proteins for researchers within the Northwestern Community (WCAS, McCormick, Feinberg) and also serves academic and industry researchers outside of Northwestern University. rPPC operates based on the two service models: (1) a Training model where Northwestern researchers use specialized bioreactor systems and participate in hands-on-training activities and (2) a Production model where staff carry out expression (mg to gm scale) and purification of recombinant or synthetic biologics, including potential therapeutic proteins and peptides, among others. Currently, the main focus of rPPC is to be a user-facility; the facility has parallel bioreactor systems for multiplexed lab-scale cultivation of microbial, insect, and mammalian cells. rPPC also serves as a production facility, providing low-cost recombinant biologics for researchers at Northwestern University.

OUR GOALS

rPPC’s goal is to transform research at Northwestern University by addressing a variety of unmet needs that can often stall cutting-edge projects. As a centralized protein production center rPPC facilitates the expression of proteins for a broad spectrum of research activities ranging from structure/function studies to producing protein therapeutic candidates such as monoclonal antibodies for in vitro and in vivo pre-clinical studies. Scalable, high-throughput cultivation enables screening mutants for phenotype studies, proteomics, stem cell characterization, media screening, strain and process optimization for the production of biochemicals, e.g. biofuels. High-throughput cell-free protein production interfaces well with HTA to “augment and expand Northwestern University’s community of multidisciplinary researchers focusing on areas of biomedical research relevant to NIH.” Protein purification completes the facility enabling a one-stop shop for researchers to clone, overexpress, and purify relevant biologics.

OUR PROFILES

The rPPC is co-directed by Michael Jewett, PhD, and Keith EJ Tyo, PhD. The co-directors bring top level expertise to implementing, building, and overseeing the direction of the institute; they are also involved in initial project planning and ensuring that projects adhere to agreed upon timelines and that deliverable deadlines are met. The day-to-day scientific operations of the rPPC are managed by Sergii Pshenychnyi, PhD. Dr. Pshenychnyi also advises on selection and design of optimal overexpression systems and optimizing protein purification. Dr. Irina Shepotinovskaya, rPPC Research Associate, provides instruments training, consultations and technical execution of the projects.

 

 

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Dr. Keith Tyo, rPPC Co-Director

Dr. Michael Jewett, rPPC Co-Director

Dr. Sergii Pshenychnyi, rPPC Managing Director

Dr. Irina Shepotinovskaya, rPPC Research Associate

 

 

 

 

 

 

OUR SERVICES

  • Protein Expression in Insect Cells

  • Protein Expression in Mammalian Cells

  • Protein Expression in Bacterial Cells

rPPC offers its services based on two service models: Training model and Production model. In accordance with the Training model, Northwestern researchers can use the rPPC specialized bioreactor systems and partake in hands-on-training activities. Consistent with the Production model the rPPC staff performs expression (mg to gm scale) and purification of recombinant or synthetic biologics including potential therapeutic proteins and peptides, among others. Please see below for a full list of our services:

TRANSFECTION/TRANSFORMATION

Electrocompetent or Chemically Competent E.coli cells are transformed by electroporation or heat shock with an expression vector carrying the target DNA, encoding the protein of interest.Transformed cells are plated to selection agar plates, glycerol stock is prepared from grown colonies.
Mammalian and Insect cells are transfected with target DNA encoding the protein of interest.

ANALYTICAL (SMALL SCALE) EXPRESSION IN E.COLI AND MAMMALIAN CELLS

The service includes small (5mL) scale expression of rProtein in E.coli and mammalian cells (CHO, HEK 293). It includes fermentation, induction and SDS-PAGE analysis.

PROTEIN EXPRESSION IN E. COLI (LARGE SCALE)

Recombinant protein expression involves the process of cultivation of transformed E.coli cells in 1L shaker flasks, 1L fermenter (up to 9 in parallel) or 5L fermenter. An additional rate is applied to expression using Q-plus and B-plus Bioreactors due to routine daily maintenance and cost of consumables (reagents, tubing, oxygen, syringes, air filters).

PROTEIN EXPRESSION IN MAMMALIAN CELL SYSTEM (LARGE SCALE)

rProtein expression involve inserting a DNA vector, encoding protein of interest, into mammalian cells (CHO, HEK293) and then culturing cells so that they transcribe and translate the desired protein.

PROTEIN EXPRESSION IN INSECT CELL SYSTEM

Insect cell cultures are transfected with wild type baculovirus and baculovirus transfer plasmid to generate recombinant baculovirus carrying a DNA of interest. Competent DH10 E.coli are transformed with baculovirus transfer plasmid to generate recombinant baculovirus carrying a DNA of interest. A PCR is performed to confirm true recombination through selection on Bluo Gal plates (Step1). Insect cells then infected with recombinant baculovirus to grow and amplify virus in higher titer (two ot three passages). The virus titer is determined by plaque assay (Step 2). During final step the cells are grown in high density and infected with amplified virus. Infected cells are cultivated and collected in 48-72 hours, followed by lysis to recover rProtein (Step 3). Charges applied depending on requested protocol steps and according to customers’ price group.

GENERATION OF MOUSE HYBRIDOMA, PRODUCING MONOCLONAL ANTIBODIES, per one fusion

Generation of hybridoma (secreting antibody) includes 2 steps: 1) Fusion of immune splenocytes and myeloma to make hybrid cells; 2) screening of newly formed hybridomas in ELISA. After testing the plates are transferred to the customer for in-lab cultivation and preservation of desirable clones.

RESCUING AND CULTIVATING EUKARYOTIC CELLS

An eukaryotic cells (mammalian, insect, hybridoma) to be rescued will be thawed, re-cultured with daily viability checking before freezing down for long time storage. Hybridoma will be tested for activity in ELISA for additional charge.

DOWNSTREAM PROCESSING OF GROWN CULTURE (BEFORE PROTEIN PURIFICATION)

The downstream processing involves cell mass separation by centrifugation, cells membrane disruption and lysis by high pressure homogenizer, followed by lysate clarification using high speed centrifugation. The unit cost is per cell mass from 1L of culture.

RECOMBINANT PROTEIN PURIFICATION

The service includes purification of rProteins from the cell lysate or culture medium using AKTAxpress chromatography system. The charge applies to one step of purification per 1L of cell culture.

TAG CLEAVAGE WITH TEV. PROTEASE

TEV protease is used to remove affinity tags from purified recombinant fusion proteins. The rProtein is incubated with TEV protease followed by separation of TEV protease and tag-less rProtein on affinity column using AKTAxpress FPLC system.

LARGE SCALE mAb PRODUCTION 

Hybridoma cells are grown in high density and culture medium (1L) is collected for subsequent antibody purification. *If the cells are grown in stationary type of bioreactor, the price applies to 50 mL unit of highly concentrated hybridoma supernatant.

DNA PLASMID PROPAGATION AND PURIFICATION

The competent E.coli cells will be transformed with the template DNA provided by customer. Transformed E.coli are cultivated and DNA mini or maxi prep are extracted. Quality control of DNA prep includes appearance on agarose gel, A260/280 ratio, concentration, quantitation by NanoDrop.

SDS-PAGE ANALYSIS

The samples from analytical or production scale of rProtein expression will be analysed by SDS-PAGE. The charge applies per 1 mini-gel.

WESTERN BLOT ANALYSIS

The service includes an analytical immune detection of a protein in the sample or cell lysates produced by rPPC upon customer request, or in the sample provided by a customer. The assay can be performed using traditional gel-blot-membrane protocol (the cost for traditional Western Blot (1 precast gel) includes labor time and consumables) or, upon customer request, by fully automated Wes Simple Western from ProteinSimple. In such a case the rate will include additionally instrument renting time (minimum 3 hours).

INSTRUMENT TIME SHARING

rPPC offers instrument time sharing on daily rates (Bioreactors – Qplus 1Lx3, Bplus 5L, High Pressure Homogenizer, AKTAxpress, Profinia FPLC, Innova-44 Incubator Shaker) and hourly rates (High Speed Centrifuge, Optima L-90 Ultracentrifuge, Synergy H1M Microplate Reader, Bio-Rad Gene Pulser XCell Electroporator, Bio-Rad T100 Thermal Cycler, NanoDrop 2000 Spectrophotometer, The Wes Simple Western Assay) based on instrument category. rPPC also offers an hourly rate for assisted instrument use.  This is in addition to the instrument use rates. It also includes instrument training time.

TECHNICAL/INSTRUMENT ASSISTANCE TIME

Technical assistance includes steps in research protocols not included in listed rPPC services (mixing the samples, aliquoting the samples, heat in oven, mixing the solutions). It also includes instrument training time, assistance in instrument set up, calibration, washing and turning off.

PROTEIN RECOVERING FROM INCLUSION BODIES

The service includes protein solubilization by denaturing agents followed by protein refolding using dialysis method. The rate applies per 1g of wet inclusion bodies pellet.

SERVICE FEES

The rPPC follows a mixed-use model and has a fee structure that bills for instrument time, specific services (i.e. analytical method development), or nuts to bolts projects. The latter scenario permits an investigator to drop off a cDNA and later receive: a) a cell line expressing recombinant protein, b) a data package describing the cell line (i.e. specific expression in pcd), purification, characterization (purity, potency, etc.), and/or c) a vial of purified protein in the amount requested for the project.

The fees for all of offered rPPC services are listed as follows:

Service

Fees

NU users

External users, CBC Open Access

External users, Academic

External users, Corporate

Transfection/Transformation (per one vector)

$175.00

$175.00

$280.00

$437.50

Analytical (Small-scale) expression in E.coli and mammalian cells

$440.00

$440.00

$704.00

$1,100.00

Recombinant Protein (rProtein) expression in E.coli/Shaker Flasks (per 1L of culture)

$380.00

$380.00

$608.00

$950.00

Recombinant Protein (rProtein) expression in E.coli/Bioreactor Bplus (per 1L of culture)

$570.00

$570.00

$912.00

$1,425.00

rProtein expression in mammalian cell system (per 1L of culture)

$3,180.00

$3,180.00

$5,088

$7,950

Protein expression in insect cell system (per 1L of culture)

$3660.00

$3660.00

$5856.00

$9150.00

Generation of Mouse Hybridoma, Producing Monoclonal Antibodies (per one fusion)

$3,160.00

$3,160.00

$5,056.00

$7,900.00

Large Scale mAb Production, Concentrated per 50ml or Non-Concentrated (per 1L of culture)

$820.00

$820.00

$1,312

$2,050.00

Downstream processing of grown culture (before rProtein purification) (per cell mass from 1L culture)

$365.00

$365.00

$584.00

$912.50

Recombinant Protein Purification (per lysate from 1L culture)

$320.00

$320.00

$512.00

$800.00

Tag Cleavage with TEV Protease
(per protein)

$520.00

$520.00

$832.00

$1,300.00

DNA Plasmid Propagation & Purification
(per plasmid)

$310.00

$310.00

$496.00

$775.00

Rescuing and Cultivating Eukaryotic Cells
(per 1 cell line)

$350.00

$350.00

$560.00

$875.00

Protein recovering from inclusion bodies
(per 1 protein)

$360.00

$360.00

$576.00

$900.00

SDS-PAGE Analysis

$165.00

$165.00

$264.00

$412.50

Western Blot Analysis

$270.00

$270.00

$432.00

$675.00

Instrument time – daily rate 1 (Innova Incubator Shakers, Yamato IC-600 Incubator)

$60.00

$60.00

$96.00

$150.00

Instrument time – daily rate 2 ( Profinia FPLC, High Pressure Homogenizer, Orbital shaker in CO2 incubator)

$70.00

$70.00

$112.00

$175.00

Instrument time – daily rate 3 (Q and Bplus Bioreactors, AKTAxpress FPLC)

$100.00

$100.00

$160.00

$250.00

Instrument time – hourly rate (High Speed Centrifuge, Ultracentrifuge, Microplate Reader, Microplate Washer, Gel Imager, NanoDrop 2000 Spectrophotometer, Ultrospec 10 Cell density meter, Gene Pulser, Thermal Cycler, TC20 Automated Cell Counter, The Wes Simple Western, iBlot2 Gel Transfer Device, iBind Western Device, Mini Gel Tanks, Biosafety Cabinet)

$25.00

$25.00

$40.00

$62.50

Technical/Instrument Assistance time – hourly rate

$90.00

$90.00

$144.00

$225.00

PROTOCOLS

  • E-COLI FERMENTATION
    E-COLI FERMENTATION

    E. coli Fermentation

  • WESTERN BLOTTING
    WESTERN BLOTTING

    Western Blot Protocol

  • STANDARD FUSION
    STANDARD FUSION

    Standard Fusion Protocol

  • CULTIVATION OF HYBRIDOMA CELLS
    CULTIVATION OF HYBRIDOMA CELLS

    Cultivation of Hybridoma Cells

  • STANDARD ELISA
    STANDARD ELISA

    Indirect Standard ELISA Protocol

INSTRUMENTS

Innova 44 Incubator Shaker

Innova 44 Incubator Shaker, New Brunswick Scientific

New Brunswick Scientific’s Innova 44 Series Lab Shakers provide large-capacity shaking for laboratories short on space. These shakers accept test tubes and flasks up to 5 liters; and can be stacked up to three units high as shown, for triple the capacity, while occupying the same space as a single shaker. Innova 44 offers incubation up to 80ºC, while Innova Model 44R adds refrigeration.

  • Superior Air Circulation System with low-watt-density resistance heaters for rapid temperature equilibration and excellent uniformity
  • Precisely-Regulated Temperature, ranging from +5ºC above ambient to 80ºC (-20ºC below ambient to 80ºC for Innova 44R); accurate within ±0.1ºC
  • Dynamic Speed Range from 25 to 400 rpm controlled within ±1 rpm
  • Timed Shaking from 0.1 to 99.9 hours, or continuous shaking
  • RS-232 Computer Interface allows remote data logging and program control using standard lab management software
  • Heavy-Duty Door Gasket maintains a leak-free seal and ensures excellent temperature uniformity
  • Extra-Large Chamber holds flasks up to 5 liters

BIOSTAT® Q plus, Screening Fermentor/Bioreactor (two instruments)

BIOSTAT® Q plus, Screening Fermentor/Bioreactor (two instruments)

The BIOSTAT® Q plus is a new generation of

Fermentor / Bioreactor systems designed for parallel operation with high throughput capability.

The combination of a newly engineered control system launches small scale cultivation vessels into a new era of biotechnology screening.

The BIOSTAT® Q plus has the capability to control fully independently up to 12 culture vessels with minimal manual operation.

For further enhancement of system performance, a powerful supervisory process control software MFCS/DA for extended visualization, data acquisition and trend display is included.

The BIOSTAT® Q plus is ideal for:

  • Process development
  • Process optimization
  • Up- and Down-scale experiments
  • Strain and Cell line characterization
  • Quality control
  • Parallel production process control

Applications:

  • Growth studies of microbial, mammalian, insect and plant cells
  • Culture media composition and optimization
  • Upscale migration, i.e. transition from shaking flasks
  • Downscale of production process for process optimization
  • Small scale protein and mAb expression
  • High cell density cultivation

Technical Specification:

  • Space requirement / weight /1 L culture vessel, 3-fold: 1100 + 900 + 800 [mm] / 114kg
  • Ambient temperature / relative humidity (non condensating) 5–40°C / 85%
  • Agitation motor speed: 50–1200 rpm
  • Temperature: 0–150 °C
  • pH: 2–12
  • pO2: 0–100%
  • Gassing System / Microbial version: Air aeration with O2 supplementation, Sparger outlet

/ Cell Culture version: 4-gasmixing for Air N2, O2, CO2, Sparger and Overlay outlet

  • Pump rotation speed: 20 rpm
  • Flow rate integrated pumps: 0.04–33.2 [ml/min] (tube dependent)
  • Thermostat temperature control range: 8°C above cooling water to 60 °C

The BIOSTAT® B plus, Fermentor/Bioreactor with O2-Enrichment Gassing System

The BIOSTAT® B plus, Fermentor/Bioreactor with O2-Enrichment Gassing System

The BIOSTAT® B plus with integrated O2- Enrichment gassing capability enables high oxygen transfer for high cell density cultures as well as sheer-stress sensitive gassing for filamentous organisms. Furthermore, it may help to solve foaming problems due to reduced gassing rates. SCADA Software BioPAT® MFCS/DA provides plug-and-play configuration, online data acquisition, Sample Data Management, enhanced plotting functions, export functions.

Application:

  • Culture of microorganisms
  • Batch, fed batch and continuous culture
  • High-cell density culture
  • Culture of filamentous microorganism
  • Small-scale cell mass and protein production
  • Anaerobic / microaeriphilic culture, on request

Key Features:

  • Single vessel configuration
  • Automatically controlled O2-Enrichment
  • Graphical user interface with touch screen operation
  • Totalizers with digital calibration for valves and pumps
  • One high-performance stirrer motor for all UniVessel® sizes
  • Trend display with up to 6 process values
  • Pre-configured firmware for system extensions

Technical Specification:

  • Space requirement / 5 L culture vessel: 620 + 730 + 565mm
  • Culture vessel: 5L
  • Agitation motor speed: 20–1,500rpm
  • Temperature: 0–150 °C
  • pH: 2–12
  • pO2: 0–100%
  • Gassing System: Air aeration with O2 supplementation, Sparger outlet
  • Gas flow range “Sparger”/ 5 L: 1.3–13 [l/min]
  • Pump rotation speed: 20 rpm
  • Flow rate integrated pumps: 0.04–33.2 [ml/min] (tube dependent)
  • Temperature control range: 8°C above cooling water to 80°C
  • Supervisory process control software: BioPAT® MFCS/DA for data storage

Avestin Emulsiflex-C5 homogenizer

Avestin Emulsiflex-C5 homogenizer

The Avestin Emulsiflex C5 is a lab scale, air driven, high pressure homogeniser that rapidly reduces particles and droplets from micron to nanometer sizes.

High pressure homogenisation is an efficient and easy to use technique for cell disruption and liposome/emulsion processing.

The principle is to pump the sample through a valve with a small aperture. The sample goes from very high pressure on the pump side of the valve to atmospheric pressure on the outlet side of the valve. This extreme change of pressure produces very high shear forces on the sample as it exits the valve. The effect on cells is to disrupt their membranes and the effect on liposomes/emulsions is that they are dramatically reduced in size.

The Emulsiflex C5 uses a dynamic homogenising valve. The benefit of a dynamic valve is that the samples are processed at a constant homogenising pressure. This produces very uniform liposomes and micro emulsions with a narrow particle size distribution.

High pressure homogenisation enables liposomes and emulsions to be processed from microns to nanometer sizes. Cell disruption is also very efficient. Most cells are ruptured in a single pass thereby avoiding over processing and protein damage.

The dynamic homogenising valve can be exchanged with a high pressure extruder. This enables the extrusion of liposomes or emulsions through membranes of specific pore sizes.

The Emulsiflex C5 requires a supply of compressed or bottled air. For most emulsions, liposomes and E. coli rupture, an air pressure of 100psi is sufficient.

Avanti® J-E Centrifuge System

Avanti® J-E Centrifuge System, Beckman-Coulter

Avanti J-E with speeds up to 21,000 rpm coupled with powerful *SR drive technology provides you with fast separations. Extensive rotor library maximizes capacity and optimizes g-force. The library includes: general purpose, high capacity (4 liter), and biosafe certified rotors with patented dual locking lids for safe sample transportation.

  • Maximum Speed: 21,000 rpm
  • Maximum g-force: 53,300 x g
  • Maximum Capacity: 4 L
  • Acceleration/Deceleration: 2/3
    • Set Temperature Range: -10°C to 40°C (1°C increments)
    • Temperature Control: 2°C of set temperature
    • Speed Control: 25 rpm of set speed or 0.1%, whichever is greater
    • Ambient Temp Range: 15°C to 35°C
    • Drive Type/Cooling: SR Drive/Air Cooled
    • Refrigeration System: Non-CFC, non-ozone depleting refrigerant
    • Dimensions HxDxW: 87 cm x 80 cm x 64 cm34 in x 31 ½ in x 25 in
    • Weight: 264 kg (583 lb.)

Optima™ L-90 K Preparative Ultracentrifuge

Optima™ L-90 K Preparative Ultracentrifuge, Beckman-Coulter

Optima™ L-90 K Ultracentrifuge generates 694,000 x g at speeds up to 90,000 rpm thus enabling to perform more separations in less time. It operates with a broad range of superb rotors, including zonal and continuous flow for large-volume separations. The L-90 K offers the reliability of a vacuum-encased induction drive, the simplicity of user-friendly, microprocessor-based control, and environmentally-friendly cooling systems.

  • Digital control via touch-sensitive keys and digital displays
  • Choice of slow-start and standard acceleration
  • Imbalance-tolerant drive
  • Set Speed: Actual rotor speed ± 20 rpm of the set speed (above 1,000 rpm)
  • Set Temperature: 0° C to 40° C in increments of 1° C
  • Set Time: Up to 99 hr and 59 min; HOLD for runs of unspecified length
  • Speed Range: 1,000 rpm to maximum speed
  • Temperature Control: ± 0.5° C of set temperature; within 0.3° C of set after equilibration
  • Door: 17.5 mm (0.69 in) high-strength structural steel
  • Humidity Restrictions: < 95% (non-condensing)
  • Time — Actual Display: Indicates time remaining in timed runs, time elapsed in Hold runs, or estimated time remaining in Ω2t runs
  • Vacuum: Diffusion pump in series with a mechanical pump typically reduces chamber pressure to below 5 µ (0.7 Pa)

Baker SterilGARD III Advance Biological Safety Cabinet

Baker SterilGARD III Advance Biological Safety Cabinet

  • 10’’ view screen sash opening
  • UV germicidal lamp
  • stainless Steel IV bar
  • Additional petcocks
  • Plastic Storage Bins
  • Air GUARD mass airflow monitor
  • Armrest
  • Ergonomic Adjustable Footrest
  • Type B3 exhaust kit, Thimble Connection
  • ·Type B3 exhaust kit, Hard Connection
  • Reinforce work Surface
  • Seismic Restraints

Cryobiological Locator 4

Barnstead/Thermolyne, Cryobiological Locator 4

 

  • Used for cold storage of biological specimens in liquid nitrogen.
  • Advanced vacuum insulation provides a maximum −190°C .
  • Delivers improved liquid nitrogen consumption rates.
  • Secure locking clasp prevents unauthorized access.
  • Each Locator vessel includes stainless steel racks with
  • Nalgene Cryoboxes for 2mL freezing vials
  • Cryoboxes are not included.
  • No of Racks: 4
  • Working Capacity (Liquid N2): 90 L
  • Static Consumption (Liquid N2): 0.65 L/day
  • Static Holding Time: 138 days
  • System Capacity: 3600 ampules
  • Height x Diameter x Diameter Neck: 914 x 559 x 216 mm

Synergy™ H1m Microplate Reader

Synergy H1m Monochromator-Based Multi-Mode Microplate Reader

Synergy™ H1m is the most cost-effective monochromator-based multi-mode microplate reader on the market. Its monochromator optics uses a third generation quadruple grating design that allows working at any excitation or emission wavelength with a 1 nm step. This system supports top and bottom fluorescence intensity, UV-visible absorbance and high performance luminescence detection. It is the ideal system for all the standard microplate applications found in life science research laboratories. Synergy H1m can be turned into a high performance Synergy H1 Hybrid Multi-Mode Microplate Reader with the addition of a filter-based optical module. A dual reagent injection system is available to automate inject/read assays such as ion channels assays or flash luminescence assays (e.g. luciferase or ATP assays).

  • Fully monochromator-based with quadruple grating architecture
  • Upgradeable to patented Hybrid monochromator-filter optical system
  • Compatible with Take3 Micro-Volume Plate: sixteen 2 µL microspots for low volume DNA/RNA 260 nm measurements. Saves precious samples, very fast and simple process.
  • High performance fluorescence (top and bottom), absorbance and luminescence detection systems accommodate all life-science research microplate applications.
  • Automated Z-height adjustment accommodates variable microplate heights and liquid levels with 100 μm resolution.
  • Optional dual-reagent dispenser can be used for assays requiring precise timing such as ion channel assays or luminescent flash ATP assays.
  • Comes with Gen5 software: reader control, advanced data analysis and flexible Excel export in one software package.

Rental of this instrument includes the usage of the attached BioTek ELx50 Microplate Strip Reader.

Profinia™ Protein Purification Automated Chromatography System

Profinia™ Protein Purification Automated Chromatography System

The Profinia protein purification system is designed for automated two-step affinity purification of histidin (His)- and GST-tagged proteins. With unattended operation, the Profinia protein purification system yields purified samples for downstream use in as little as 30 minutes. The instruments takes up minimal bench space by integrating all reagent bottles, fraction tubes, sample vials, and tubing into a closed, compact system. The Profinia instrument displays details for each significant step, including expected run times, to allow you to track run progress. Full run data are captured as purification occurs, then are calculated, displayed, and stored. Using a USB flash drive, run data can easily be exported. For real-time data acquisition, the instrument can be connected to a PC loaded with optional Profinia software.

Purification data are displayed on your PC monitor in real time, and can be stored, analyzed, and customized for specific reporting needs.

Instrument specifications:

  • Flow rate range: .2–20 ml/min
  • Maximum backpressure : 45 psi/3.4 bar
  • Operating temperature range: 4–30ºC
  • UV absorbance wavelength: 280 nm
  • Conductivity monitor range: 0–500 ms/cm
  • pH monitor (optional) pH range: 1–14
  • Hardware : Sample and fraction 15 or 50 ml conical tubes; can use larger external reservoirs connected by tubing
  • Collection containers: Buffer selection rotary selector valves for up to 8 application-specific buffers
  • Solvent compatibility: All commonly used chromatographic solvents
  • Reporting software: Windows 2000 or XP; chromatographic data display, reporting, and analysis in real time or using stored run files
  • User interface: Touch screen 115 x 86 mm;
  • Dimensions (W x D x H): 58 x 33 x 67 cm

NanoDrop 2000, Micro-Volume UV-Vis Spectrophotometer

NanoDrop 2000, Micro-Volume UV-Vis Spectrophotometer

The Thermo Scientific NanoDrop 2000 is the only
micro-volume spectrophotometer with a patented sample retention technology that allows for sample volumes as small as 0.5 µL. NanoDropTM 2000 Spectrophotometer enables fast and highly accurate UV/Visible analyses of samples with remarkable reproducibility. Spectrophotometer ideally suited for such applications as purified protein and nucleic acid concentration and purity, cell density measurements, Bradford and BCA protein assay.

  • Direct, easy measurements in less than 5 seconds – just pipette & wipe
  • Full spectral output
  • Measures DNA, RNA (A260) and Protein (A280) concentrations and sample purity (260/280 ratio)
  • Large concentration range (2 ng/µL – 15,000 ng/µL dsDNA) without dilutions
  • Pre-configured methods for common applications such as Nucleic Acid, Protein A280, Microarray, Proteins & Labels, Bradford, BCA, Lowry , and more
  • User-friendly software that includes Custom Methods and data export capabilities
  • Simple self calibration check using control fluid – no instrument adjustments
  • Low-cost operation – no plates or other consumables

Gene Pulser Xcell™ Total System

Gene Pulser Xcell™ Total System

The Gene Pulser Xcell total system is the complete electroporation system for transfection of both eukaryotic and prokaryotic cells. The Gene Pulser Xcell total electroporation system includes the main unit, the capacitance extender, or CE, the pulse controller, or PC module, ShockPod™ cuvette chamber, 15 sterile electroporation cuvettes (5 each of 0.1, 0.2, and 0.4 cm electrode gap), and cuvette rack.

Key Features and Benefits

  • Universal electroporator capable of transfecting all cell types
  • Compatible with any electroporation buffer, including Bio-Rad’s Gene Pulser® electroporation buffer
  • Preset protocols for most popular mammalian and bacterial cell types
  • Flexible manual programming
  • Extensive collection of transfection protocols

Gel Doc™ EZ System

Gel Doc™ EZ System

  • Ease of use — researchers not familiar with the imager can use it easily because there is no need to manually control filters, lens, or lights. Users can create default protocols once and then use the green button on the front of the instrument to reproducibly use these settings time and again
  • Delivery of publication-quality images and completely analyzed results — get clean, smooth, publication-ready images with decreased pixelation when images are cropped or zoomed. Analyzed results, including relative molecular weights, quantitation of bands, Excel reports, PDFs, and more are available in a matter of minutes
  • Time and space savings — obtain results faster without spending time on steps normally required for image acquisition. Save valuable benchspace using this compact system
  • Modular design and flexible options — use specific trays for specific applications. The clearly defined, color-coded trays eliminate confusion. Purchase only what you want and upgrade when your needs change
  • Stain-free technology — condense your 2 hr Coomassie protocol into a 5 min stain-and-image step with this stain-free imaging system. Stain-free gels are western blot compatible, allowing you to check electrophoresis results and quality prior to western blotting. These “green” gels provide sensitivity equal to or better than Coomassie staining and eliminate organic waste disposal concerns

TC20™ Automated Cell Counter

TC20™ Automated Cell Counter

The TC20 automated cell counter counts mammalian cells in one simple step using its innovative auto-focus technology and sophisticated cell counting algorithm to produce accurate cell counts in less than 30 seconds. Upon insertion of a counting slide, the TC20 cell counter rapidly provides a total cell count (with or without trypan blue staining) and assesses cell viability via trypan blue exclusion.

  • Compatible with a broad range of cell sizes and types — counts cell lines, primary cells (from tissue or blood), and stem cells
  • Innovative auto-focus technology — removes the variation associated with manual focusing and leads to precise cell counts in 30 seconds
  • Cell size gates — user selects a population of interest in complex samples, such as primary cells, or lets the cell counting algorithm do all the work
  • Cell viability — analyzes cells accurately usingmultifocal plane analysis
  • Easy to archive and analyze — stores up to 100 counts in the onboard memory for access any time, or use the optionalTC20 data analyzer software on your PC to further analyze exported cell images

AKTAxpress Chromatography System

AKTAxpress, GE Healthcare

ÄKTAxpress™ is a robust and easy-to-use system for multistep purification of affinity-tagged proteins and antibodies. Purpose-designed hardware, intelligent software, and pre-packed media enable high protein purity and ensure maximum productivity. ÄKTAxpress for tagged protein purification gives you the highest possible purity needed for structural and functional studies. Optimized protocols with a choice of up to four purification steps minimize the need for chromatography expertise. Tag removal and maintenance procedures can be integrated into the purification protocols, eliminating manual interference during a run. Purification schemes for double affinity-tagged proteins are also supported. The system is capable of purifying target proteins in yields varying from 1 to 50 mg or more. Small footprint – a compact design allows the system to occupy minimal space on the bench or in the cold room.

System specifications:

• System flow rate 0.1 – 65 ml/min
• Operating pressure 3 MPa (30 bar, 435 psi)
• UV wavelength range 280 or 254 nm
• pH range 0 – 14 (spec. valid between 2 and 12)
• Conductivity range 1 μS/cm to 999.9 mS/cm
• Solvent compatibility – all commonly used chromatographic solvents
• Module Size 250 mm × 650 mm × 700 mm (W × D × H)
• Weight 27 kg/module

Invitrogen™ Mini Gel Tank

Invitrogen™ Mini Gel Tank

The Mini Gel Tank is compatible with a variety of Novex® mini gels, NuPAGE® mini gels, and Bolt® Bis-Tris Plus gels. Each Mini Gel Tank can accommodate up to two gels per run. The unique tank design enables convenient side-by-side gel loading and enhanced viewing during use. Optimized conditions using constant voltage allow for Bolt® Bis-Tris Plus gels to be run in approximately 35 minutes. Run times may vary depending on buffer conditions and power supplies being used.

The Mini Gel Tank offers:

  • Versatility—compatible with NuPAGE™, Novex™, and Bolt mini gels
  • Easy sample loading—forward-facing well configuration
  • Simultaneous visualization of both gels—streamlined, side-by-side tank configuration
  • Simple monitoring of prestained protein markers— with white tank stand
  • Less running buffer required—two separate gel chambers, so you only need to load sufficient buffer for each gel to the specified fill line
  • High performance—same great results you’ve been getting with the XCell SureLock™ Mini-Cell and original Bolt Mini Gel Tank

The Wes Simple Western

The Wes Simple Western

The Wes is the instrument that makes running fully automated Western blot, or Simple Western size assay. The Wes employs a novel fully automated Capillary Electrophoresis (CE)-based immunodetection technology, called the Simple Western size assay. CE is a technique designed to separate ionic species based on their size in the interior of a small capillary filled with electrolytes. This technology is based on nanovolume size-based protein separation that can be used to quantify proteomic profiles of any experimental protein samples, or clinical specimens for both biomarker discovery and diagnostics. Instead of two days you spend just three hours to get accurately quantitated, and reproducible result of 25 samples!

Assay Principles: A Simple Western by size is an automated Western—no gels, no transfer devices, no blots, no film and no manual analysis. Just load your samples in Wes and press start—it’s a complete, walk-away solution for protein detection.  Wes automate all steps automatically including sample loading, size-based  protein separation, immunoprobing, washing, detection and data analysis.

Variability in the manual processes that used to impact reproducibility, quantitation, time to result and overall data reliability is eliminated. Transition is simple—the same antibodies used for Western blotting protocols can be used with Simple Western assays. Up to 25 samples can be processed at once, and assays take just 3 hours to complete with sizing and quantitation results.

The assay kits come with a pre-filled plate that contains all the reagents other than the researcher’s samples and primary antibodies. Samples for Wes are separated in a self-contained capillary cartridge, and immobilized to the capillary wall via a proprietary UV capture method. Target proteins are immunoprobed with an antibody followed by HRP-amplified chemiluminescent detection. Wes automates the entire Western blot procedure increasing reproducibility and delivering significant workflow time savings.

Simple Western size-based assay data is processed automatically in Compass software. Sample data is displayed by lane in a virtual-blot like image similar to traditional results with one big exception—not only do you get more information, you get it as soon as the assay is complete. Quantitative result such as molecular weight, signal intensity (area), % area, and signal-to-noise for each immunodetected protein are presented in the results table automatically.

iBlot™ 2 Gel Transfer Device

iBlot™ 2 Gel Transfer Device

The iBlot® 2 Gel Transfer Device is our second-generation dry transfer device, providing the same speed and convenience as the original iBlot® device, but with many new features. The iBlot® 2 Gel Transfer Device performs western blotting transfer simply, efficiently, and reliably, within seven minutes and without the need for liquid buffers. The iBlot® 2 Gel Transfer Device is an integral part of the iBlot® 2 Dry Blotting System, which consists of the transfer device and consumable transfer stacks that contain the required buffers and transfer membrane (nitrocellulose or PVDF). The iBlot® 2 device and the transfer stacks are sold separately.

Features of the iBlot® 2 Dry Blotting System:

• Complete protein transfer in 7 minutes or less
• High detection sensitivity and even transfer
• Increased blotting reliability and reproducibility
• Flexible gel size formats and membrane types
• A simple, user-friendly system
• Options to create new custom programs
• Built-in tutorial and application notes
• High-quality transfer stacks that are more compact than before

Using the iBlot® 2 System for Rapid, High-quality Protein Transfers
The iBlot® 2 Dry Blotting System is for dry electroblotting of proteins from mini-, midi-, and E-PAGE™ gels onto nitrocellulose or PVDF membranes for western detection. The iBlot® 2 system offers high-quality transfer, convenience, and speed, producing crisp, clear bands that remain sharp and straight, with exceptional transfer efficiency. Additionally, the iBlot® 2 Dry Blotting System offers new options that allow users to create their own custom programs.

How It Works
Buffer ion reservoirs are incorporated into the gel matrix (transfer stacks) instead of buffer tanks or soaked papers. The high density of ions in the gel matrix enables rapid protein transfer. During blotting, the copper anode does not generate oxygen gas as a result of water electrolysis, reducing blot distortion (conventional protein transfer techniques, including wet, semi-wet, and semi-dry, use inert electrodes that generate oxygen). With the iBlot® 2 Dry Blotting System, transfer time is reduced by the shortened distance between electrodes, high field strength, and high current. Trapped air bubbles, often created during the manual preparation of the blotting sandwich layers, are easy to avoid due to our unique de-bubbling design that promotes even and complete transfer.

iBlot® 2 Transfers Save You Time
With the iBlot® 2 system, there is no need to prepare buffers, pre-treat your gel, or clean up after blotting. The total preparation and run time is normally less than 10 minutes per blot. Compared with conventional western transfer techniques, iBlot® 2 transfers can save you hours.

iBind™ Western Device

iBind™ Western Device

The iBind™ Western Device is an automated western-processing device that performs every step from blocking to washes to antibody incubations via sequential lateral flow (SLF). SLF allows the timely release and flow of solutions and antibodies to the membrane without need of an external power source. Each solution is released from the iBind™ device wells to an iBind™ Card via SLF; the solutions are then wicked towards the stack region of the card. The glass fiber matrix of the card allows for homogenous and consistent flow of the solutions to the membrane, increasing the antigen-antibody interaction.

Features of the iBind™ Western Device include:

• Simple, automated solution
• No external power source
• Superior reproducibility over traditional manual methods
• Better antigen antibody interaction—solutions flow between the iBind™ Card and membrane for maximum interaction between protein and antibody

The iBind™ western detection procedure minimizes hands-on time and is highly reproducible and compatible with all western protocols. Set up time is fast: within 2.5 h you are ready for the detection step.

 

 

 

CONTACT US

 

CONTACT INFORMATION

Dr. Sergii Pshenychnyi
Managing Director
Recombinant Protein Production Core at CLP
Northwestern University

847.467.5303
sergii.pshenychnyi@northwestern.edu

VISIT US

Chemistry of Life Processes Institute

Silverman Hall, Rm. B715
2170 Campus Drive
Evanston, IL 60208-2850


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