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GE Healthcare

Capture the perfect image for winning resultsQuantitative imaging solutions

Capture the perfect image for winning resultsGE Healthcare offers you a wide variety of high-quality products for 1-D and 2-D electrophoresis, Western blotting, and 2-D Difference Gel Electrophoresis (DIGE) workflows. Our extensive range of electrophoresis and blotting instruments and reagents plus our imaging, labeling and detection solutions, provide you with the highest quality results for the identification and quantitation of proteins of interest.

Look in depth and uncover detailsGE Healthcare’s imaging, labeling and detection solutions give you a clearer view. Our range of high-performance, versatile, and upgradeable imagers provide a comprehensive system for every experimental need. We have also developed powerful software for integrated high-quality image analysis.

Amersham ECL™ Western Blotting Detection Reagents were the world’s first commercial chemiluminescent reagents for Western blotting and they are still the most widely used. They are at least ten times more sensitive than colorimetric assays and give reliable results time after time. We offer reagents with different sensitivities for a wide range of applications.

The ECL Plex™ fluorescent Western Blotting system offers multiplexing possibilities for reliable quantitation of your target protein relative to a reference protein.

A proven reputation in Western blotting and 2-D DIGEOur Western blotting solutions are renowned for delivering reproducible results of the highest quality. Products like ECL Western Blotting Detection Reagents and Hybond™ membranes are among the most cited for their applications in peer-reviewed publications. The Ettan™ DIGE System for differential analysis of complex protein samples has generated over 1000 peer-reviewed publications since its introduction to the scientific community in 1997.

End-to-end solutions, validated to work togetherTo help you attain the highest quality of quantitative results from biomolecular studies, GE Healthcare provides validated end-to-end solutions. We take full responsibility for the entire detection and analysis technical workflow, from protein and nucleic acid sample preparation, through electrophoresis, blotting, labeling, image acquisition, and data analysis. By providing uncompromising quality in every step of the process, we help you derive the most information from your experiments.

Capture the perfect image and achieve the quantitative results your applications demand. Add labeling, detection and imaging solutions from GE Healthcare to your winning team today.

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High quality in the entire biomolecule detection and analysis workflow ensures optimal quantitation

Electrophoresis Analysis

SDS-PAGE, IEFMini format – Vertical

miniVE

SE 250 / SE 260

EPS 301

Purification kitsTrap Protein Sample Preparation

Sample Grinding KitIgG and Albumin Removal KitSDS-PAGE Clean-Up Kit2-D Clean-Up KitMini Dialysis KitVivaspin concentrators

PlusOne Reagents

FluorescenceECL Plex

CyDye™ Antibody Labeling Kits

Fluorescent markers and stains

ECL Plex Fluorescent Rainbow Markers

Deep Purple™ Total Protein Stain

Quantitative detectionTyphoon™

Storm™

Ettan DIGE Imager

ImageQuant™ RT ECL

ImageQuant 350

ImageQuant 300

ImageScanner III

Qualitative detectionImageQuant 150

Hyperfilm™

SoftwareImageQuant TL

ImageMaster™ 2D Platinum

DeCyder™ 2D

DeCyder EDA

DeCyder MS

Chemiluminescent markers

ECL DualVue™ Markers

ECL Western Blotting Molecular Weight Markers

Chemiluminescent and chemifluorescent reagents

ECL HRP-linked 2º Abs/streptavidin

ECL Protein Phosphorylation and Biotinylation Modules

ECL GST and Glycoprotein Detection Modules

ECF™ Western blotting reagent

ChemiluminescenceECL Advance

ECL Plus

ECL

ProcessingProcessor Plus

ECL Multiprobe

BlottersminiVE blot module

TE 22

MembranesHybond ECL

Hybond-P

Hybond-LFP

BlottersTE 62

ECL Semidry Blotter TE 70 / TE 77 / TE 70 PWR / TE 77 PWR

Standard formatSE 600 Ruby

EPS 601

Automated – Flatbed

PhastSystem

2-DFirst dimensionEttan IPGphor 3

Immobiline DryStrip gels

Second dimensionMini – miniVE

Standard – SE 600 Ruby

Large – Ettan DALTsix, Ettan DALTtwelve

Protein

Spectrophotometers & kitsNanoVue

GeneQuant 1300

2D Quant Kit

Amplification kitsillustra nucleic acid amplification kits

Purification kitsillustra nucleic acid purification kits

NonradioactiveAlkPhos Direct with CDP-Star or ECF detection

ECL Direct Nucleic Acid Labeling

Gene Images Random-Prime Labeling

Gene Images 3’-Oligolabeling

SpectrophotometersNanoVue

GeneQuant 1300

BlottersVacuGene XL

TE 22, 42, 62, with EPS 2A200

MembranesHybond-N+

Hybond-NX

Hybond-N

Hybond-XL (radioactive only)

Hybond Blotting Paper

SystemsUVC 500 Crosslinker

Hybridization Oven

MacroVue

RadioactiveRedivue nucleotides

Rediprime II

Rapid-hyb Buffer

Ready-To-Go DNA Labeling Beads

5’-End Labeling

Nick Translation

MegaPrime

CDP-Star™ (chemiluminescence)

ImageQuant 350

ImageQuant RT ECL

Hyperfilm ECL

Hypercassette

ECL (enhanced chemiluminescence)

ImageQuant RT ECL

ImageQuant 350

Hyperfilm ECL

Hypercassette

Hyperprocessor

ECL Plus & ECF (enhanced chemifluorescence)

Typhoon

Storm

QuantitationTyphoon

Storm

SoftwareImageQuant TL

AnalyticalSmall < 1000 bpPolyacrylamide - GenePhor

Medium 100 bp to 10 kbMini – Agarose - HE33

EPS 301 Power Supply

Standard – Agarose - HE99X units

EPS 301 Power Supply

Large 10 kb to 10 MbPulsed field - Gene Navigator

Gel mobility assayCy™3 and Cy5

Nucleic acid

DetectionLabelingBlottingQuantitationSample Preparation

www.gelifesciences.com/sampleprep www.gelifesciences.com/nanovue www.gelifesciences.com/2DE www.gelifesciences.com/western www.gelifesciences.com/quantitative_imaging

Electrophoresis AnalysisDetectionLabelingBlottingQuantitationSample Preparation

General purpose film RadioactiveHyperfilm MP

Hypercassette

2-D DIGE

CyDye DIGE Fluor Minimal Kit

CyDye DIGE Fluor Kit for Scarce Samples

Chemifluorescence

ECL Plus

ECF Substrate

Typhoon variable mode imagersSuperior resolution, versatility, and sensitivity for all imaging applicationsThe ability to distinguish subtle expression differences among biomolecules present in low amounts in complex mixtures is vital for genuine biological discovery. GE Healthcare provides versatile tools for pioneering scientific research—solutions that offer superior versatility for the selection of an optimal imaging approach, linearity of signal response, quantitative accuracy, and extremely low limits of detection.

When measuring subtle changes, the true measure of quality in any analytical instrument is the signal-to-noise ratio. By combining powerful lasers (generating high signal), confocal optics (to focus on the true signal), and industry leading twin Hamamatsu™ photomultiplier tubes (for the lowest noise or interference in signal detection), the Typhoon Variable Mode Imager is the ultimate standard for image quality.

The versatility of Typhoon imagers enables you to select the most suitable imaging technique for a particular experiment. By choosing Typhoon, researchers step outside the bounds set by limited imaging platforms, gain versatility for optimal imaging and uncover the biology to reveal phenomena that they might otherwise miss.

• Large format imaging systems with highest imaging versatility: filmless phosphor imaging, multiplex fluorescence, and chemiluminescence

• Five to one hundred times faster than film

• Excellent linear dynamic range with five orders of magnitude

• Optimized signal-to-noise providing excellent limit of detection

• Reusable storage phosphor screens

• Fully optimized as part of the Ettan DIGE System

• Supplied with leading ImageQuant™ TL analysis software

The Typhoon Variable Mode Imager offers:

• Fully optimized and automated multicolor scanning that allows for the detection of multiple samples in the same image. This ensures accuracy of analysis, increases throughput, and saves time

• Multiple sample formats—gels, blots, sandwich gels, phosphor screens, microplates, and plate assays

• Storage phosphor, fluorescence, and chemiluminescence modes in one instrument

• High-power laser excitation and ultra-low noise

• Unique scanning head mechanism ensures targeted, fast but powerful imaging capability with minimal loss of signal through photo bleaching of the fluorophor

• Method driven selection of filters to ensure optimal performance

• Simultaneous scanning of two large 2-D gels on the sample area of 35 x 43 cm (14 x 17 in).

The basis for signal detection in analytical instrumentation

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Typhoon versatility

The versality of the Typhoon Variable Mode Imager has been demonstrated for a variety of applications including:

• Northern and Southern blotting

• Chemifluorescent and fluorescent Western blotting

• 2-D DIGE

• Analysis of PCR products

• Hybridisation studies

• Post-translational modification studies

• Fluorescence in microtiter plates

• Enzyme assays

• Dot blots

• Detection of DNA-protein complexes

• Tissue selections

• Radioactive metabolic labeling

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Intensity plots of a dilution series show background, signal, and noise levels during detection. Typhoon instrument ensures high signal and low noise, to detect even the lowest signals above prevailing background and noise

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Relative protein quantitation with ECL Plex

ECL Plex Western blotting system works with sensitive CyDye conjugated antibodies and low fluorescent membranes for the multiplexed detection of both low- and high-abundant proteins simultaneously.

ECL Plex Western blotting system allows for the detection of low-abundant proteins with high linearity and a dynamic range of 4 orders of magnitude using Typhoon. By using multiplexed ECL Plex fluorescent Western blotting, it is possible to simultaneously detect two target proteins on the same blot by using CyDye conjugated antibodies. Therefore, reliable quantitation relative to a housekeeping protein can be easily obtained. This is important to avoid false conclusions due to experimental variation.

Specific multiplex detection of ß-Tubulin and ERK 1/2 using ECL Plex Western blotting system and Typhoon

Sample: CHO cell lysate, four-fold dilutions, 5 µg to 78 pg

Membrane: Hybond LFP

Primary antibodies: mouse anti-tubulin and rabbit anti-MAP kinase ERK1/2

Secondary antibodies: ECL Plex anti-rabbit Cy5, ECL Plex anti-mouse Cy3, ECL Plex anti-rabbit Cy3, ECL Plex anti-mouse Cy5.

Typhoon

Matrix type: membrane

Pixel size: 100 µm

Channels: Cy3 Cy5

CHO cell lysate (µg)

5 1.25

0.31

0.07

8

5 1.25

0.31

0.07

8

Cy3 channel

Cy5 channel

tubulin

ERK 1/2

ECL Plex Cy5 and Cy3 attain the same sensitivity (dye-independent, giving true relative levels) and they can detect concentrations as low as 78 pg total protein in lysate.

Multiplexed detection of two proteins on the same blot enables reliable quantification of the protein of interest (POI) relative to a housekeeping protein (HKP), eliminating experimental variation such as uneven sample loading. This image shows p38 activation upon TGF-stimulation for 0, 2.5, 5 and 15 minutes in 293T cells. Phosphorylated p38 was detected using ECL Plex Cy5 and the housekeeping protein actin was detected using ECL Plex Cy3.

Housekeeping protein (HKP)

Protein of interest (POI)

Ratio (POI/HKP)

B

N

Phosphor imaging

• Typhoon imagers can be used to quantitate a broad range of isotopes, including 3H, 14C, 125I, 18F, 32P, 33P, 35S and 99mTc

• Simultaneously detects strong and weak signals within five orders of magnitude linear dynamic range

• Compatible with most available storage phosphor screens

• Reusable storage phosphor screens and image eraser are available from GE Healthcare, saving time and cost compared to using film

• Typically five times faster compared to using film

Relative quantitative differences of protein levels in healthy and cancerous tissue from the same patients with diagnosed early stage (T1/T2) colorectal cancer was studied using the Ettan DIGE system minimal labeling approach, where all samples are normalized to an internal standard. Of 288 unique proteins identified using MALDI-TOF MS, 30 were found to be differentially regulated by 2-D DIGE and Typhoon. Only 15 of the differentially regulated proteins had previously been reported to be involved in cancer.

Cy3—Normal Cy5—Cancer

Stathmin

PEKAM1

MRP14

eEF -

Muscle fructose 1.6 bisphosphate Aldolase

Normal Tumor

Tropomyosin 1

Tropomyosin 2 ß

Ser proteinase inhibitor Clade A

ACTB (actin ß)

Creatine kinase

Normal Tumor

The Typhoon imager continues to be used in pioneering and innovative research—your imagination combined with a versatile, sensitive, and robust imaging platform. Some examples of recent novel applications using Typhoon include:

• Quantitative multiplexed fluorescence Western blotting (ECL Plex)—identify and quantitate 2 target proteins on the same blot without stripping and reprobing

• Tissue section imaging

• Cell surface protein labeling—identify and quantitate signaling proteins expressed on the cell surface

• Blue-Native DIGE—separation and quantitation of intact protein complexes

• Quantitative profiling of membrane proteins from laser capture microdissection

• Analysis of scarce samples—quantitative analysis of samples less than 1000 cells (0.5 µg of protein)

• DIGE combined with 16-BAC and SDS-PAGE for the analysis of hydrophobic proteins

• 3-Dimensional profiling of plasma and serum using 2-D LC, 1-D electrophoresis and DIGE labeling

• Biomarker discovery, phosphorylation, and protein modification studies using 2-D DIGE

• Measuring the redox potential of proteins using 2-D DIGE

• Plant ozone damage, environmental stress responses, and photosynthesis using 2-D DIGE

Highly selective labeling of cell surface proteins using intact cells dramatically increase the possibility of detecting these low abundant proteins compared to the standard Ettan DIGE labeling protocol.

Quantitation of potential biomarkers for colorectal cancer by 2-D DIGE

Standard Ettan DIGE protocol

Mix

2-D DIGE

Cy3Total lysate

Cy5Cell lysate

Cell surface protocol

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Storm imaging systems High performance phosphor imaging and mono and dual fluor applications on a proven platformStorm gel and blot imaging systems are the economical laboratory standard in variable mode imagers. The systems offer a wider dynamic range than film and accurate signal quantitation that yield publication-quality images on first exposure. Storm systems use storage phosphor screens to deliver high-resolution imaging and accurate quantitation of 14C, 3H, 125I, 32P, 33P, 35S, and other sources of ionizing radiation. Choose from three different models, each one with proven capability for phosphor imaging (autoradiography and radioisotope detection). Storm also provides different types of fluorescence imaging for nucleic acid and protein gel analyses; these systems also offer chemifluorescence imaging for blot analyses within minutes—eliminating the need for film. Storm imaging systems offer intuitive operation that is independent of the application. Simply place your gel, blot, or storage phosphor screen on the glass platen; then point-and-click on the scan control window to start your scan. ImageQuant TL image analysis software, included with every Storm system, provides powerful data analysis and reporting capabilities.

• Large format imaging systems for filmless phosphor imaging, single and dual fluorescence, and chemifluorescence imaging

• Five to one hundred times faster than film

• Excellent linear dynamic range with over five orders of magnitude

• Reusable storage phosphor screens and image eraser

Sensitive, linear detection response

This application shows a two-fold dilution series showing the excellent linearity and high sensitivity of ECL Plex. CyDye conjugated secondary antibodies detect proteins up to three orders of magnitude. Multiplex fluorescent Western blotting is possible with Storm using ECL Plex Cy5 and Cy2.

Single channel chemifluorescence

Fast blot analysis Chemifluorescence is an easy-to-use technique for nonradioactive DNA, RNA, and protein detection on blots. With chemifluorescence, there is no need for multiple film exposures because the Storm system reads your blots in minutes. Further, quantitative analysis is simplified because a digital image is acquired directly, removing the need to scan an exposed film.

Easy, familiar and stable protocols Chemifluorescence sample preparation is very similar to chemiluminescence sample procedures. The advantage is that the fluorescent reaction product is stable and does not diminish over time and allows accurate quantitation, which gives reproducible and accurate results between blots and enables the running of a larger number of blots in a single experiment.

Sample: human apo-transferrin, two-fold dilutions, 5 ng to 0.6 pg

Membrane: Hybond LFP

Primary antibodies: rabbit anti-human transferrin

Secondary antibodies: ECL Plex goat anti-rabbit Cy2

Storm system

Pixel size: 100 µm

Channel: Cy2 (Laser: 450 nm, Em. Filter 520 nm LP)

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Phosphor imaging with Storm

Storm PhosphorImager uses storage phosphor screens instead of film to capture data from radioactive gels and blots, which provide superior sensitivity compared to film. A choice of screens to detect different isotopes is available. Screens are sensitive to any source of ionizing radiation, including commonly used isotopes such as 32P, 33P, 35S, 14C, 3H, and 125I. Storm has a large 35 x 43 cm (14 x 17 in) sample area. With its excellent dynamic range, Storm captures the image from both strong and weak signals in a single exposure. The linear dynamic range is 1000 times greater than film and produces images on storage phosphor screens five to one hundred times faster compared to film.

Storage phosphor screens are available in several formats and are durable and long-lasting. With care, storage phosphor screens last indefinitely, regardless of how often they are used. A ten-minute exposure to a visible light image eraser (delivered with each Storm) prepares the screen for reuse.

Macroarray imaging of small or large format cDNA array is easily performed for gene expression profiling.

Courtesy of Christopher Hug, Dept. of Cell

Biology, Washington University Medical School,

St. Louis, Missouri, U.S.A.

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DR: 101.8 (5ng to 78 pg)LOD: 78 pg

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Transferrin (ng)

DR = Dynamic RangeLOD = Limit Of Detection

Ettan DIGE ImagerDedicated fluorescence imager for 2-D DIGEEttan DIGE Imager is based on innovative scanning technology for fluorescence imaging with a novel, progressive scanning CCD system, which eliminates the disadvantages of widefield CCD imaging. It is suitable for a wide range of fluorescent gel or blot imaging applications, especially 2-D DIGE experiments.

This user-friendly imager offers high dynamic range, excellent resolution and sensitivity for the accurate detection of the faintest features. These specifications are crucial for effective imaging of 2-D electrophoresis gels, particularly for acquiring multiplexed images of Cy2-, Cy3-, and Cy5-labeled gels or blots.

Ettan DIGE Imager is supplied with comprehensive, but intuitive image acquisition software with the flexibility to image a wide variety of gel formats and fluorophores.

• High resolution imaging from a novel progressive scanning CCD that eliminates CCD array distortion and pixel variability, thereby contributing to more accurate quantitation than widefield CCD imaging

• Optimized and balanced fluorescence detection in multiplexed experiments, cutting experimental bias to a minimum

• High sensitivity for imaging faint spots combined with 16-bit image data that allows over 3.5 orders of magnitude in linear dynamic range for accurate and simultaneous quantitation of low and high concentrations

• Dedicated, removable sample handling cassettes for naked gels, membranes, DALT and SE 600 series gel formats

• Advanced low fluorescent materials in the optics path and sample cassette keep noise to a minimum. Shielded optics inhibit sample heating and fluorophor degradation during scanning

• Powerful inexpensive excitation source that is easily replaced by the user

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Ettan DIGE Imager

Matrix type: gel

Pixel size: 100 µm

Channels: Deep Purple 1 (Ex. Filter: 540/25 nm, Em. Filter: 595/25 nm) SYPRO Ruby 1 (Ex. Filter: 480/30 nm, Em. Filter: 595/25nm)

Analysis software: ImageQuant TL

Sample: Carbonic anhydrase, two-fold dilutions, 1 µg to 0.06 ng

Protein stains: Deep Purple, SYPRO™ Ruby

Detection of Deep Purple and SYPRO Ruby total protein stains with Ettan DIGE Imager

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Measuring differential protein abundance

Ettan DIGE system together with Ettan DIGE Imager and DeCyder 2D Differential Analysis software is a powerful tool for accurately quantitating changes in protein abundance. CyDye DIGE Fluor minimal dyes enable multiplexing of three separate protein samples on one second-dimension SDS PAGE. Multiplexing with 2-D DIGE allows the incorporation of the same internal standard on every gel to eliminate gel-to-gel variation.

The benefits of 2-D DIGE for quantitation combined with the dynamic range and linearity of Ettan DIGE Imager enables you to measure low expression and high and low abundant proteins in the same gel—helping you to go beyond the limits of proteomics to visualize and accurately quantitate the most subtle of protein expression changes. By generating overlaid, multichannel images for each gel, Ettan DIGE Imager integrates the ability to multiplex and ensures seamless data handling throughout the DIGE analysis process. Images are co-detected and analyzed using the DeCyder Differential Analysis software to accurately measure very small protein differences with high confidence. Since its introduction to the scientific community in 1997, Ettan DIGE system has generated over 1000 peer-reviewed publications, making GE Healthcare the company of choice for providing end-to-

Image of a 2-D DIGE gel from Ettan DIGE Imager showing excellent image correspondence

Ettan DIGE system workflow (from Westermeier R, Scheibe B. Difference gel electrophoresis based on lys/cys tagging. In Posch A. Ed. Sample Preparation

for 2D PAGE. Methods in Molecular Biology, Humana Press, Totowa, NJ (2008) 73-85).

Labeled proteinsSamples Co-migration in 2-D electrophoresis

Image analysis

IPG strip

SDS gel

mixScan at different

wavelengths

Cy3 image

Cy5 image

Cy2 image

pooled internal standard

control

treated

label with Cy3

label with Cy5

label with Cy2

end application support from sample preparation, 2-D electrophoresis, blotting, labeling, image acquisition and analysis. As the developers of the Ettan DIGE technology platform, we continue to innovate and invest in new applications of this exciting technology.

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DR = Dynamic RangeLOD = Limit Of Detection

ImageQuant imager rangeVersatile chemiluminescence, monochromatic fluorescence, and colorimetric imagersImageQuant Imagers are flexible systems that cover a range of monochromatic imaging applications including chemiluminescence, fluorescence, and gel documentation. These CCD-based imagers provide an affordable option for protein and DNA detection with genuine benefits, including better quantitation than film without sacrificing sensitivity, publication grade data, and image and data archiving. As your needs evolve, most instruments can be upgraded from entry level systems to more advanced, high performance models.

The camera cabinet is common to most instruments and replicates a darkroom environment that includes UV and white light sources. This can be combined with epi-illumination light options which enable the imaging of, for example, silver, Coomassie and Ethidium bromide stained gels, blots, colonies (colony counting), and film. Further, the ImageQuant Imager range is delivered with powerful ImageQuant TL software, so if you already have a scanner from GE Healthcare, you won’t need to learn new software.

• Systems range from simple publication-ready gel documentation to highly sensitive detection capabilities

• Colorimetric, fluorescent, and chemiluminescent applications on a single platform

• All the advantages of film detection, with the added benefits of publication-ready images, digital archiving, and a higher dynamic range for better quantitation

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ImageQuant 300

ImageQuant 300 is a high resolution system for general gel documentation in larger multi-user laboratories.

• Fluorescence and densitometry applications, for documentation and quantitation

• High-specification cabinet with multiple lighting options

• Less risk of saturated images—10-bit camera for wider dynamic range

ImageQuant 350

ImageQuant 350 extends the performance of ImageQuant 300 for gel documentation, fluorescence detection, and chemiluminescence, thus providing higher quality data for quantitation.

• For chemiluminescence, fluorescence, and densitometry

• 16-bit, 2 megapixel camera gives dynamic range for quantitation

• Designed to support quantitative analysis of chemilumines-cent Western blots with a sensitivity that is equal to that of film. ImageQuant 350 exceeds the linearity and dynamic range of film without the saturation of strong bands.

• Eliminates the need to work in the darkroom and provides images that are ready for quantitation, annotation, sharing, and publication

• Can be switched in seconds from imaging DNA and RNA gels to Western and nucleic acid blots to 1-D and 2-D protein gels to dot blots and culture dishes

• A choice of motorized or manual lenses and emission filter supports multiple applications

• True 16-bit cooled camera enables the most accurate and cost-efficient quantitation for its price

ImageQuant RT ECL

ImageQuant RT ECL is well suited for applications that stretch the limits of sensitivity such as limited antibody or sample.

When combined with Amersham ECL Advance, the system addresses the most demanding chemiluminescent Western blotting applications.

• For chemiluminescence, fluorescence, and densitometry

• Highest sensitivity 16-bit, 4 megapixel camera for the most demanding samples

• Real-time refresh rate provides ease of use and rapid image capture

• Combinations of lens, camera, and appropriate Amersham ECL reagents deliver excellent chemiluminescent imaging sensitivity with fast exposure times

• Inherently low-noise camera with operating temperature independent of ambient temperature ensures low back-ground, high performance, and consistency

• Easy operation with control from the cabinet directly or via a computer with optional motorized lens

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ImageQuant 150

ImageQuant 150 creates high resolution records for archiving or publishing. Economical and easy to use, it is small enough to fit on your desk but powerful enough to record your electrophoresis gel and blotting membrane results. The system is optimized to capture fluorescence images without the need for film.

• Fluorescence and densitometry applications, for documentation without quantitation

• High resolution 10 megapixel , 8-bit camera

• Imaging without film or computer saves time and money

Colorimetric detection

Fluorescence detection

Chemiluminescence detection

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Film (Exp: 5 min)

Detection of chemifluorescent and fluorescent protein stains

ImageQuant RT ECL offers the highest sensitivity and resolution with minimal noise and is thereby the optimal choice for applications requiring quantitative data analysis detecting very low abundant proteins.

STAIN EXCITATION SOURCE EMISSION FILTER

Deep Purple UV transilluminator 365 nm 595/50 nm

SYPRO Ruby UV transilluminator 302 nm 595/50 nm

Coomassie Fluor Orange UV transilluminator 365 nm 595/50 nm

Sample: Carbonic anhydrase, two-fold dilutions, 250 ng to 0.24 ng

Stain: Deep Purple total protein stain, SYPRO Ruby, Coomassie Fluor Orange

Lens: Zoom lens, 28 to 70mm, F2.8

Analysis software: ImageQuant TL

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Both ImageQuant 350 and ImageQuant RT ECL imagers offer a genuine alternative to film-based detection methods. ImageQuant 350 is particularly suited to the quantitative analysis of Western blots and fully compatible with the ECL range of detection reagents. In a comparative study of an ECL Plus Western Blot, ImageQuant 350 achieved similar sensitivity (limit of detection) to film-based detection, but with a significant improvement in dynamic range. The signal response for ImageQuant 350 was linear over the entire range of protein concentration.

ImageQuant 350 FilmMatrix typ

Exposure time: 6 min 5 min

Lens: Fix lens, 28 mm, F1.8

Analysis software: ImageQuant TL

Sample: human apo-transferrin two-fold dilutions, 5 ng to 9.8 pg

Membrane: Hybond ECL

Pixel size: 100 µm

Detection reagent: ECL plus

Comparison of ImageQuant 350 with film for ECL Plus chemiluminescence detection

ImageQuant 350

DR: 102.7

(5 µg to 9.8 pg)

LOD: 9.8 pg

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Hyperfilm ECL

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Saturated bands

Quantitative Western blotting

Sensitivity is an important feature for an imager, especially when your experiment involves the detection of a protein expressed at low levels. Also, a broad linear dynamic range is a requirement for applications comparing weak and strong signals in the same image. Limitations in dynamic range of some imaging systems can, for these types of applications, result in incomplete datasets and limited results.

ImageQuant 350 and ImageQuant RT ECL imagers meet both of these requirements. By offering a broad dynamic range, 16-bit cooled CCD, as well as high sensitivity, your chances of a successful experiment are improved.

For chemiluminescence detection, the use of film can provide good sensitivity. However, the drawback of using film is the limited dynamic range. To obtain high sensitivity using film, increased exposure times are required resulting in saturated signal from proteins of high concentration. ImageQuant 350 and ImageQuant RT ECL provide the same level of sensitivity as film with a wider dynamic range and this leads to the quantitation of both strong and weak bands in the same gel.

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)

Transferrin (pg)

24 25

Dynamic range

Dynamic range

5000 2500 1250 625 310 156 78 39 19.5 9.8 pg

250

ng

125

ng

62.5

ng

31.3

ng

15.6

ng

7.8

ng

3.9

ng

1.95

ng

0.98

ng

0.49

ng

0.24

ng

C Coomassie Fluor Orange

DR: 102.4 (250 ng to 0.98 ng)LOD: 0.98 ng carbonic anhydrase

50

40

30

20

10

00 50 100 150 200 250 300In

tegr

ated

inte

nsity

(x10

-4)

Carbonic anhydrase (ng)

ImageQuant RT ECL

R2 = 0.9871

B SYPRO Ruby

60

50

40

30

20

10

00 50 100 150 200 250 300In

tegr

ated

inte

nsity

(x10

-4 )

Carbonic anhydrase (ng)

ImageQuant RT ECL

R2 = 0.985

A Deep Purple

30

25

20

15

10

5

00 50 100 150 200 250 300In

tegr

ated

inte

nsity

(x10

-4)

Carbonic anhydrase (ng)

ImageQuant RT ECL

R2 = 0.9986

C Coomassie Fluor Orange

Three fluorescence-based gel protein stains; Deep Purple, SYPRO Ruby and Coomassie Fluor Orange were imaged using ImageQuant RT ECL. In this experiment, Deep Purple shows the best sensitivity and the highest dynamic range.

ECL Exposure time: 20 min

DR: 102.1 LOD: 39 pg transferrin

ECL Advance Exposure time: 1 min 30 sec

DR: 102.4 LOD: 19.5 pg transferrin

R2 = 0.9952

R2 = 0.99646

Film offers a sensitive, linear response at low concentrations (lower graph), but its dynamic range is limited compared to ImageQuant 350 (upper graph).

DR = Dynamic RangeLOD = Limit Of Detection

DR = Dynamic RangeLOD = Limit Of Detection

ImageScanner IIIVersatile densitometer with high linearityImageScanner III

Versatile densitometer with high linearity

ImageScanner™ III is a versatile, high resolution scanner with an exceptional optical density range. It offers the accuracy and sensitivity essential for the precise quantitation for a wide range of samples. For example, it can perform imaging of classical 1-D and 2-D gels (non-DIGE) stained with Coomassie or silver.

• Variable scanning rate for faster image acquisition prevents gel drying

• Full 16-bit pixel depth for accurate quantitation of weak and intense image features

• Selectable color for better sensitivity and reduced background

• Sealed environment for scanning wet samples and modified transmission to accommodate gels between glass plates

• High sensitivity for capturing faint sample features without requiring a diffusion plate

• Samples can be scanned in either reflection or transmission mode

Quantitative analysis of proteins in 2-D

Scanning Coomassie or silver 2-D gels using ImageScanner III and further analyzing with ImageMaster 2D Platinum software enables detection of biological variation in the order of 2-fold with very low risk of false positive or negative results.

ImageScanner III is well suited for quantitative analysis of proteins in 1-D because of its large linear range and consistency of scanning.

ImageScanner III is calibrated with a linearity above 4.0 OD which means that stains will be the limiting factor and not the instrument. When ImageScanner III is used with a step–calibration tablet and LabScan™ 6.0 software, linearity and quantitation is greater than 3.6 OD. The scanning light has the option to drop out colors depending on the stain used, which means that silver stained gels can be scanned with blue color to provide higher quality images.

ImageScanner III shows a high degree of consistency in spot or band detection and quantitation. After repeated scanning of the same image and subsequent image analysis, the average CV (Coefficient of Variation) of spot or band volumes was typically less than 5% for over 90% of features. ImageScanner III in conjunction with ImageMaster 2D Platinum or ImageQuantTL software enables accurate detection and quantitation of proteins with a high degree of confidence.

Densitometry and quantitative analysis of proteins in 1-D

28 29

Image analysisFrom images to real results

Our software products address a wide range of image analysis needs—from basic documentation and purity screening to the querying of entire gene expression or 2-D gel datasets. Combined with our imaging instruments, our software makes a complete system to provide total solutions to many challenging applications.

1-D, Array, Colony counting—ImageQuant TL

• Manual or automated, easy-to-use image analysis software

• Four functional areas for 1-D, dot blots, colony counting/2-D spot measurement, and user-defi ned gel analysis

2-D—ImageMaster 2D Platinum

• Analysis of 2-D DIGE and non-DIGE 2-D electrophoresis gels

• A fl exible and intuitive package that gives you control over your analysis

2-D DIGE—DeCyder 2D

• Dedicated software for 2-D DIGE analysis, employing state-of-the-art algorithms

• Spots real differences with statistical confi dence

• Time-saving automation saves days of work

Advanced statistics—DeCyder EDA

• Advanced statistical analysis for DIGE data output from DeCyder 2D

• Calculates, interprets, and reports—using multivariate analysis to uncover patterns in expression data

ImageQuant TL

DeCyder 2D

ImageMaster 2D Platinum

DeCyder EDA

DeCyder 2D Differential Analysis Software

Ettan DIGE system eliminates gel-to-gel variation in 2-D electrophoresis to reveal real biological variation. The system is based on the specific properties of CyDye DIGE Fluor dyes which enable multiplexing protein samples on the same 2-D gel, and the inclusion of an internal standard.

DeCyder 2D software automatically locates and analyzes multiple samples in a gel and then enables comparative analysis of multiple gels, producing accurate measurement of differential changes in protein abundance.

• Complete 2-D DIGE analysis including patented co-detection algorithms

• Internal standards derive statistical data within and between gels to eliminate variation

• Detection of as low as 10% differences in protein levels with statistical confidence

• Reduction of time for hands-on analysis to minutes

DeCyder Extended Data Analysis Software

DeCyder Extended Data Analysis Software (DeCyder EDA) is an add-on module to DeCyder 2D with powerful, high- performance informatics software developed to complement the 2-D DIGE methodology. As a high-performance proteomics informatics software, DeCyder EDA was developed specifically for 2-D DIGE and therefore all the advantages of the DIGE approach are utilized in the software analyzing 2-D DIGE gels to easily create pick lists of specific proteins.

EDA is used for multivariate analysis of protein expression data and can handle up to 1000 spot maps. Raw data (gel images) linked to EDA can undergo a range of statistical analyses and automated interpretation. These include:

• Principal component analysis Produces an overview of the data. Can be used to find outliers in the data

• Pattern analysis Finds patterns in expression data (e.g. proteins and spot maps with similar expression profiles)

• Discriminant analysis Finds proteins that discriminate between different samples (to find biological markers), creates classifiers and assigns samples to known classes depending on expression profiles (e.g. tumor typing)

• Interpretation Finds the biological context of proteins by integrating biological information and context from in-house or public databases. It can be used to determine in what pathways and processes a protein is involved, the function of the protein etc.

ImageQuant TL Software—automated 1-D and 2-D analysis

ImageQuant TL is a suite of image analysis tools designed to meet a broad range of analysis requirements. ImageQuant TL is an automated and easy-to-use general purpose get, blot and array image analysis software.

• Intuitive user interface with step-by-step interactive guides

• High levels of automation as well as full manual control for detection and quantitation

• Comprehensive editing functions at each stage of the analysis

• Automatic report builder with full user control for laboratory notebooks and publication

• Analysis of multiplexed images

It offers four separate functional areas that include: 1-D electrophoresis gels; dot blots, microplates, and basic arrays; colony counting and measurement of 2-D spots; and a toolbox to analyze gels with user-defined objects.

The flexible ImageQuant TL software features high levels of automation, yet still allows manual editing and overrides at any stage of the analysis. Image data can be extracted easily and accurately in a matter of seconds to produce consistently reliable data. ImageQuant TL software also supports multiplexing for advanced fluorescence applications. ImageQuant SecurITy adds functions for audit trails and authorization for GMP compliant laboratories.

ImageMaster 2D Platinum v6.0 user interface with dockable windows simplifies the process of 2-D gel image analysis.

ImageMaster 2D Platinum Software— flexible 2-D analysis

ImageMaster 2D Platinum is built for comprehensive visualization, exploration, and analysis of 2-D gel data. Developed by the Swiss Institute of Bioinformatics in collaboration with GeneBio and GE Healthcare, ImageMaster 2D Platinum is powered by the widely used Melanie gel analysis software. It simplifies and improves the analyses of 2-D gels and the identification of protein markers of interest.

• Established methods for all 2-D experiments, including single stain and DIGE

• Easy import and image viewer functionality

• Effective spot detection and matching solutions

• Versatile analytical methods

• User-friendly and flexible interface

• Adaptable visualization tools

• Seamless integration in your laboratory workflow

• High quality and integrity of data

ImageMaster 2D Platinum can also analyze 2-D DIGE experiments. It uses the patented co-detection algorithm of DeCyder™ 2D software to detect differences between samples as low as 50%.

32 33

Single-click identification of gel lanes and bands. DeCyder 2D Differential In-gel Analysis facilities comparison of samples by linking the user interface data inspection quadrants

A gene ontology query in DeCyder EDA.

Amersham Hyperfilm ECL compared with Kodak BioMax Light

Increased sensitivity and superior contrast

Amersham Hyperfilm MP compared with Kodak BioMax MS

Excellent performance without a premium priceAmersham HyperfilmAmersham Hyperfilm ECL: The clear choice for reliable qualitative chemiluminescent detectionApplications: chemiluminescent detection in Western, Southern, and Northern blots

• Increased sensitivity: greater sensitivity for low concentration protein detection

• Clearer background: superior contrast and band visibility in the detection of chemiluminescent signals

• Artifact-free results: minimal occurrence of visual artifacts due to the improved anti-static layer

• Validated performance: compatibility with ECL, ECL Plus, ECL Advance™, CDP-Star and all blue and green chemiluminesence systems

35

Applications: 32P, 33P, 125I Northern, Southern, and Western blots 35S, 33P, 32P sequencing Fluorography of 3H, 14C, and 35S

• High sensitivity: sensitivity down to a single-copy gene detection

• Clearer results: a clearer, colorless background that improves band visibility

• Flexibility: compatibility for use with all commonly used radioisotopes

Amersham Hyperfilm MP: The clearest results in radioactive detection

Comparative use of Kodak BioMax MS and Amersham Hyperfilm MP in dot blot experiments using 32P (400, 200, 100, 50, 25, 12.5, 6.25, 3.125, 1.56, 0.78 dpm) with an exposure time of 24 h.

Kodak BioMax Light

Amersham Hyperfilm ECL

Western blot detection with Kodak™ BioMax™ Light and Amersham Hyperfilm ECL films. Performance of New Hyperfilm ECL and Kodak BioMax Light with Western blots of immobilized IgG from mouse protein (5 µg, 2.5 µg, 1.25 µg, 635 ng, 310 ng, 155 ng, 77 ng, and 39 ng). Detected with horse radish peroxi-dase labeled anti-mouse antibody 1:5000 and Amersham ECL Western Blot detection reagents.

Amersham Hyperfilm ECL compared with Kodak BioMax MS

Increased sensitivity

Kodak BioMax MS

Amersham Hyperfilm ECL

Western blot analysis with Kodak BioMax MS and Amersham Hyperfilm ECL films using rabbit IgG (155, 75, 37.5, 18.75, 9.37, 4.7, 2.34, 1.17, 0.58, 0.3 ng) and Amersham ECL Advance detection reagents 5 min exposure.

dpm/band

400

200

100

50

25

12.5

6.25

3.125

1.56

0.78

BioMax MSAmersham Hyperfilm MP

GE, imagination at work and GE monogram are trademarks of General Electric Company.

Amersham, Cy, DeCyder, Deep Purple, ECL, ECL Advance, ECL Plex, Ettan, Gene Navigator, GenePhor, GeneQuant, Hybond, Hyperfilm, ImageMaster, ImageScanner, ImageQuant, MacroVue LabScan, NanoVue, Storm, Typhoon and VacuGene are trademarks of GE Healthcare companies.

2-D DIGE: 2-D Fluorescence Difference Gel Electrophoresis (2-D DIGE) technology is covered by US patent numbers 6,043,025, 6,127,134 and 6,426,190 and equivalent patents and patent applications in other countries and exclusively licensed from Carnegie Mellon University.

ImageMaster 6: This version of ImageMaster has been developed by the Swiss Institute of Bioinformatics in collaboration with GeneBio and GE Healthcare

DeCyder: This release of DeCyder version 2.0 (software) is provided by GE Healthcare to the customer under a non-exclusive license and is subject to terms and conditions set out in the 2-D Differential Gel Electrophoresis Technology Access Agreement. Customer has no rights to copy or duplicate or amend the Software without the prior written approval of GE Healthcare.

For contact information for your local office, please visit: www.gelifesciences.com/contact

GE Healthcare Bio-Sciences ABBjörkgatan 30751 84 UppsalaSweden

www.gelifesciences.com/quantitative_imaging

Deep Purple Total Protein Stain: Deep Purple Total Protein Stain is exclusively licensed to GE Healthcare from Fluorotechnics Pty Ltd. Deep Purple Total Protein Stain may only be used for applications in life science research. Deep Purple is covered under a granted patent in New Zealand entitled “Fluorescent Compounds”, patent number 522291 and equivalent patents and patent applications in other countries.

ECL Advance: ECL Advance contains Lumigen TMA-6 sub-strate and is sold under exclusive license from Lumigen Inc.

CyDye: This product or portions thereof is manufactured under an exclusive license from Carnegie Mellon Universityunder US patent number 5,268,486 and equivalent patents in the US and other countries. Cy3-UTP or Cy5-UTP, Cy3. 5-dCTP or Cy5.5-dCTP, Cy3-CTP or Cy5-CTP: These products are manufactured for GE Healthcare UK Limited by Perkin Elmer Life Sciences under US patent numbers 5047519 and 5151507. The cyanine dyes in the product are manufactured under an exclusive license from Carnegie Mellon University under US patent numbers 5,268,486 and equivalent patents in the US and other countries.The purchase of CyDye products includes a limited license to use the CyDye products for internal research anddevelopment but not for any commercial purposes. A license to use the CyDye products for commercial

purposes is subject to a separate license agreement with GE Healthcare. Commercial use shall include: 1. Sale, lease, license or other transfer of the material

or any material derived or produced from it. 2. Sale, lease, license or other grant of rights to use this

material or any material derived or produced from it. 3. Use of this material to perform services for a fee

for third parties, including contract research and drug screening.

If you require a commercial license to use this material and do not have one, return this material unopened toGE Healthcare Bio-Sciences AB, Bjorkgatan 30, SE-751 84 Uppsala, Sweden and any money paid for the materialwill be refunded.

Coomassie is a trademark of ICI plc. SYPRO is a trademark of Invitrogen Corp. CDP-Star is a trademark of Tropix Inc. Hamamatsu is a trademark of Hamamatsu Photonics.Kodak and BioMax are trademarks of Eastman Kodak Co.

©2008 General Electric Company—All rights reserved. First published August 2008.

GE Healthcare UK LimitedAmersham PlaceLittle ChalfontBuckinghamshire, HP7 9NAUK

GE Healthcare Europe, GmbHMunzinger Strasse 5D-79111 FreiburgGermany

GE Healthcare Bio-Sciences Corp.800 Centennial Avenue, P.O. Box 1327Piscataway, NJ 08855-1327USA

GE Healthcare Bio-Sciences KKSanken Bldg., 3-25-1, HyakuninchoShinjuku-ku, Tokyo 169-0073Japan

28-9419-67 AA 08/2008