ChemiDoc™ MP Brochure - Bio-Rad

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Imaging
ChemiDoc™ MP Imaging System
Confidence in every step of
your imaging workflow.
ChemiDoc MP
Imaging System
Because results always matter.
Bring a new level of capability and efficiency to your
experiments with an imaging system designed for
multitasking. The ChemiDoc MP Imager is a unique
imaging system that enables stain-free operation
and lets you visualize proteins at every stage of your
blotting experiment. Its flexibility and sensitivity are
complemented by simple, intuitive operation that
integrates seamlessly into your workflow.
bio-rad.com
Superior Sensitivity
Get quantitative, reproducible data without relying on outmoded film
processes. The ChemiDoc MP Imaging System offers advanced detection
technology that creates optimal exposure for even the faintest bands.
Rely on it for fast, super-sensitive chemiluminescence and fluorescence
detection and for colorimetric gel and blot documentation.
Signal-to-noise ratio of transferrin. 10 sec exposure.
180.0
170
160.0
140.0
Signal-to-noise ratio
Fig. 1. Sensitivity
comparison of the
ChemiDoc MP System
versus X-ray film
using blots of serial
dilution of transferrin.
A, the ChemiDoc Imager
delivers superior dynamic
range and comparable
limit of detection to film.
B, a 10-second exposure
on film reveals a more
limited dynamic range
than the ChemiDoc MP
System. Saturated pixels
are highlighted in red.
131
121
120.0
114
95
100.0
93
85
80
60
60
40
48
30
29
28
28
27
27
26
20
0
30
20
15
ChemiDoc MP Imaging System
10
7.5
5.0
Film
Fig. 1B
Film
Sample load, ng
ChemiDoc MP Imaging System
24
3.8
2.5
Sample load, ng
Fig. 1A
Bio-Rad Laboratories, Inc.
58
47
34
23
26
22
1.9
1.3
0.63
32
19
19
0.31 0.15
17 14
0.08
SUPERIOR SENSITIVITY
Fig. 2. A calibrated
luminescent target
was used to test light
collection efficiency
and determine overall
system sensitivity.
A, data from lower
limits of detection are
graphed to show overall
signal-to-noise ratio. The
ChemiDoc MP System
delivers top-of-the-class
chemiluminescence
sensitivity against leading
multiplexing imagers on
the market. B, images of
lower limits of detection
from a calibrated
luminescent target. The
ChemiDoc MP System
delivers excellent image
quality and limit
of detection.
Signal-to-noise ratio of LEDs from a calibrated luminescent test plate. 5 min exposure.
100
90
90.7 89.8
80
Signal-to-noise ratio
70
60
50
40
30
20
22.1 21.6
20.3 20.2
10
5.9
6.6
6.2
7.5
1.9
2.0
3.6
4.1
0
0.7
0.9
12
34
LED well
ChemiDoc MP Imaging System
Competitor 1
Competitor 2
Competitor 3
Fig. 2A
Fig. 2B
ChemiDoc MP
Competitor 2
12 34
Competitor 1
12 34
Competitor 3
ChemiDoc MP Imaging System
bio-rad.com
Exceptional Image Quality
With patented focus calibration technology, images are always in focus at
any zoom level. Exceptional dynamic range enables visualization of faint and
intense bands on same blot or gel. With Image Lab™ Software you can edit
and analyze images on the spot without exporting to other programs.
Fig. 3. 1-D
Coomassie-stained gel.
Fig. 4. Chemiluminescent
ELISA arrays. Quansys
Biosciences Q-Plex Array
has 16 distinct capture
antibodies bound to each
well of a 96-well plate.
Fig. 5. Fluorescent
multiplex blot with
DyLight 488, DyLight
549, and DyLight 649
conjugates.
Fig. 3
Fig. 5
Bio-Rad Laboratories, Inc.
Fig. 4
EXCEPTIONAL IMAGE QUALITY
Minimal cross-talk between blue and green channels.
Fluors detected in green channel
Cross-talk
Correct
Fluors detected in blue channel
Correct
Cross-talk
FAM
A488
Cy3
A546
FAM
A488
Cy3
A546
2.3
1.1%
100%
100%
100%
100%
0.1%
0.0%
FAM
A488
Cy3
A546
FAM
A488
Cy3
A546
12.4%
7.6%
100%
100%
100%
100%
0.2%
0.0%
FAM
A488
Cy3
A546
FAM
A488
Cy3
A546
32.5%
21.3%
100%
100%
100%
100%
9.7%
6.1%
ChemiDoc MP
Competitor 1
Fig. 6A. Determination
of cross-talk using
same multiplex gel.
Blue fluors (FAM and
Alexa Fluor 488) are
detected in the green
channel and green fluors
(Cy3 and Alexa Fluor
546) in the blue channel
using the ChemiDoc
MP System and a
competitor’s system.
For competitor 1, the
bold percentage values
indicate high cross-talk.
Fig. 6A
Competitor 2
Fig. 6B
Cross-talk
FAM
A488
Correct
Cy3
A546
Correct
FAM
A488
Fig. 6B. Visualization
of cross-talk signals
recorded with a
second competitor’s
instrument. The green
and blue channel
images show high
cross-talk values
and are indicated in
bold. The blue fluors
(FAM and Alexa
Fluor 488) are detected
in the green channel
and the green fluors
(Cy3 and Alexa Fluor
546) are detected in
the blue channel.
Cross-talk
Cy3
A546
ChemiDoc MP Imaging System
bio-rad.com
Unmatched Application Versatility
The ChemiDoc MP Imager is the only system you need
when your experiments include a variety of sample types or
require different detection methods. It is the perfect imager to
accompany your protein and DNA electrophoresis runs as well
as your western blotting experiments. And it delivers quantitative,
reproducible results every time.
Fig. 7. Multiplex image
file allows multichannel
and individual channel
view. Image of multiplexed
fluorescent blot with
Alexa Fluor 488, Alexa
Fluor 555, and Alexa
Fluor 647.
Bio-Rad Laboratories, Inc.
UNMATCHED APPLICATION VERSATILITY
Fig. 8. Multiple
applications of the
ChemiDoc MP Imager.
DIGE Criterion™ Gel
EtBr-stained wide
mini ReadyAgarose™ Gel
GelRed-stained
mini ReadyAgarose Gel
Silver-stained 2-D Criterion Gel
Coomassie-stained
Mini-PROTEAN® TGX™ Gel
SYPRO Ruby-stained
large-format 2-D gel
Flamingo™-stained
Mini-PROTEAN TGX Gel
Criterion Stain Free™ TGX Gel
SYBR® Green-stained wide mini
ReadyAgarose Gel
ChemiDoc MP Imaging System
bio-rad.com
Ease of Use
The ChemiDoc MP System is designed for productivity. With little or
no training, users can acquire publication-quality images in seconds.
The system is precalibrated to provide the precise focus for any
zoom setting or sample height; automated hands-free operation
ensures consistent, reproducible, and high-throughput performance.
Fig. 9A. The application selected automatically determines the excitation sources and emission filters.
Fig. 9B. The ChemiDoc MP System is always in focus at any zoom level — no more out-of-focus gel
or blot images.
Bio-Rad Laboratories, Inc.
EASE OF USE
Autosetup
Autofocus
Autoexposure
Autooverlay
Autoanalysis
Autoreports
Autoprint
ChemiDoc MP Imaging System
bio-rad.com
Stain-Free Enabled
Stain-free technology, offered only by Bio-Rad, eliminates extra steps and
unproductive delays in your western blotting experiments.
Stain-free technology is the keystone of the Bio-Rad® V3 Western Workflow™,
a portfolio of products that enables researchers to visualize, verify, and validate
results at each step of their western blotting experiment.
V3
Stain-free gel
Stain-free blot
Total protein quantification
Protein of interest probed with Alexa Fluor 649
Visualize
The ChemiDoc MP System will activate
and provide immediate visualization
of protein separation in all lanes with
stain-free gels before blot transfer.
Bio-Rad Laboratories, Inc.
Verify
The ChemiDoc MP System paired
with stain-free technology enables
instant verification of protein transfer
before blot detection.
Validate
Flexible Image Lab Software tools
normalize for protein of interest using
total protein stain from stain-free blot
and provide quantitative blot results.
STAIN-FREE ENABLED
V3 Western Workflow
The V3 Western Workflow streamlines the western blotting protocol, incorporating stain-free in-gel chemistry to allow rapid fluorescent
detection of proteins for gels and blots as well as the use of total protein normalization as a loading control. This improved workflow saves
time and increases accuracy and reliability throughout the western blotting process.
Workflow
Benefit
Separate Proteins
Run gels in as little as 15 min
Speed with flexibility: TGX Stain-Free™ Gel chemistry available in precast and handcast formats
1
■■
Visualize Protein Separation
Visualize separation for all lanes in 1 min
2
■■ Coomassie-like performance with no background variability and no staining/destaining
Stain-free image of
pretransferred gel
Transfer
Efficient and uniform protein transfer
in as little as 3 min
3
■■
Throughput: transfer 4 mini gels at once
Verify Transfer Efficiency
Quickly assess transfer efficiency
4
■■
Verify quality of transfer for all lanes in 2 min
Stain-free image
of blot
Antibody Incubation and Blot Detection ~5 hr
Validate Western Blot Data by
Normalization and Analysis
Use stain-free blot image as total protein
loading control
■■
5
No need to strip and reprobe
Use the entire protein sample in one lane
(no need to rely on housekeeping proteins)
■■
Detect protein
of interest
Normalize protein
of interest with
stain-free image of
blot from step 4
■■
Reliable and accurate quantitation
ChemiDoc MP Imaging System
bio-rad.com
Total Protein Normalization
Stain-free gel chemistry makes it possible to use total protein levels as a loading control rather than the housekeeping proteins
used in traditional western blotting protocols. This negates the need to strip and reprobe the blot and avoids the attendant errors
that can be introduced in this step.
1
Relative signal intensity of protein bands
Using total protein normalization
produces a much greater linear dynamic
range for measuring target protein levels.
Housekeeping proteins such as b-actin,
b-tubulin, or GAPDH are often very
abundant in biological samples, which
results in their signal being oversaturated
compared to target proteins. Normalizing
results to a total protein measurement
corrects this problem, allowing a
meaningful comparison even with
low-abundance targets, and leads to
far greater quantitative accuracy in
measuring proteins of interest.
MORE RELIABLE to quantitate the protein load
6
Stain-free total protein versus
housekeeping proteins
5
b-actin
GAPDH
4
b-tubulin
Quantitative response
3
2
1
0
0
2
Linearity comparison
of stain-free total
protein measurement
and immunodetection
of three housekeeping
proteins in 10–50 µg of
HeLa cell lysate.
Stain-free
10
20
30
40
HeLa lysate load, µg
50
60
AVOID FALSE FINDINGS caused by housekeeping protein signal saturation
50
HeLa lysate load, µg
40
30
20
False Findings: Kinase Protein Levels
Normalized against Actin
10
2.5
Akt
Erk
b-actin
Signal intensity (artificial units)
MEK
2
1.5
1
0.5
0
10
HeLa lysate load, µg
40
Truth Revealed: Kinase Protein Levels
Normalized against Total Protein
Stain-free
blot image
Signal intensity (artificial units)
2.5
2
1.5
1
0.5
0
10
HeLa lysate load, µg
Bio-Rad Laboratories, Inc.
40
Comparison of kinase
protein levels normalized
by stain-free total protein
or actin loading controls.
10–50 µg of HeLa cell
lysate from the same
sample were loaded onto
a stain-free gel to probe
MEK ( ■ ), Akt ( ■ ), and
Erk ( ■ ). There should be
no changes in the kinase
protein levels when data
are normalized. Expected
value ( ).
TOTAL PROTEIN NORMALIZATION
Research Papers Using Stain-Free Technology as
Loading Controls in Western Blots
Methodology: How stain-free total protein normalization improves western blotting
Stain-free total protein staining is a superior loading control to b-actin for western blots.
Gilda JE, Gomes AV (2013). Anal Biochem 440, 186–188.
V3 stain-free workflow for a practical, convenient, and reliable total protein loading control in
Western blotting.
Posch A, Kohn J, Oh K, Hammond M, Liu N (2013). J Vis Exp, video ID 50948.
Cancer Biology
BRCA2 is epistatic to the RAD51 paralogs in response to DNA damage.
Jensen RB, Ozes A, Kim T, Estep A, Kowalczykowski SC (2013). DNA Repair (Amst) 12, 306–311.
Differential network analysis applied to preoperative breast cancer chemotherapy response.
Warsow G, Struckmann S, Kerkhoff C, Reimer T, Engel N, Fuellen G (2013). PLoS One, article ID 0081784.
Neurobiology
Proteomic analysis of gliosomes from mouse brain: identification and investigation of glial
membrane proteins.
Carney KE, Milanese M, van Nierop P, Li KW, Oliet SH, Smit AB, Bonanno G, Verheijen MH (2014).
J Proteome Res [published online ahead of print Nov 4, 2014]. Accessed Nov 10, 2014.
Hippocampal extracellular matrix levels and stochasticity in synaptic protein expression increase with age
and are associated with age-dependent cognitive decline.
Vegh MJ, Rausell A, Loos M, Heldring CM, Jurkowski W, van Nierop P, Paliukovich I, Li KW, Del Sol A, Smit AB,
Spijker S, van Kesteren RE (2014). Mol Cell Proteomics 13, 2975–2985.
A trifluoromethyl analogue of celecoxib exerts beneficial effects in neuroinflammation.
Penta AD, Chiba A, Alloza I, Wyssenbach A, Yamamura T, Villoslada P, Miyake S, Vandenbroeck K (2013).
PLoS One 8, e83119.
Skeletal Muscle
Identification of the immunoproteasome as a novel regulator of skeletal muscle differentiation.
Cui Z, Hwang SM, Gomes AV (2014). Mol Cell Biol 34, 96–109.
Immunology
Activation of the P2X7 receptor induces the rapid shedding of CD23 from human and murine B cells.
Pupovac A, Geraghty NJ, Watson D, Sluyter R (2014). Immunol Cell Biol [published online ahead of print
Aug 26, 2014]. Accessed Nov 10, 2014.
Metabolomics
Quantification of protein copy number in yeast: The NAD+ metabolome.
Mei SC and Brenner C (2014). PLoS One 9, e106496.
ChemiDoc MP Imaging System
bio-rad.com
Instrument Tour
Optimized emission and excitation filters for discrete
detection of multiplexed visible fluorescence.
Red
Automatic detection of red fluorescence
using dyes such as Cy5, Cy5.5,
Alexa Fluor 647, Alexa Fluor 680,
DyLight 649, DyLight 680, IRDye 680.
Green
Automatic detection of green fluorescence
using dyes such as Cy3, Flamingo,
Krypton, Pro-Q Diamond, Alexa Fluor 546,
DyLight 549, Rhodamine.
Blue
Automatic detection of blue fluorescence
using dyes such as Cy2, Coomassie Fluor
Orange, Alexa Fluor 488, DyLight 488,
Pro-Q Emerald 488, Qdot 523, Qdot 605,
Qdot 625, Qdot 705.
Bio-Rad Laboratories, Inc.
INSTRUMENT TOUR
Cooled CCD with autofocus
for all zoom levels
Unmatched sensitivity
and image quality
Multiplexed fluorescent imaging
Multicolor LEDs optimized for
quantitative western blot imaging
Sample tray with UV,
white, and blue imaging
Flexible transillumination
source for easy and uniform
imaging of gels and blots
6-position automated
filter wheel
Broad application flexibility
Touch-pad operation
For positioning and
band excision
Go to bio-rad.com/chemidocmp
to see the complete ChemiDoc MP video.
ChemiDoc MP Imaging System
Specifications
Ordering Information
Automation Capabilities
Workflow automated selectionApplication driven; user selected or
recalled by a protocol
Catalog #
Workflow automated executionControlled by a protocol via
application-specific setup for
image area, illumination source, filter,
analysis, and reporting
Workflow reproducibility100% repeatability via recallable
protocols; from image capture to
quantitative analysis and reports
Autofocus (patent pending)Precalibrated focus for any
zoom setting or sample height
Image flat fielding* Dynamic; precalibrated and
optimized for every application
Autoexposure2 user-defined modes
(intense or faint bands)
Hardware specifications
Maximum sample size (L x W)
28 x 36 cm
Maximum image area (L x W)
26 x 35 cm
Maximum image area for
25 x 26 cm
standard UV-excited gels (L x W)
Excitation sourceTrans-UV (302 nm included; 254 nm
and 365 nm available as options)
and epi-white; optional trans-white
conversion screen, XcitaBlue™
Conversion Screen, and epi-red,
-green, and -blue LED modules
Illumination control8 modes available. Trans-UV,
epi-white, and no illumination for
chemiluminescence are standard;
epi-red, epi-green, epi-blue, transwhite, and XcitaBlue Conversion
Screens are optional
Description
170-8280
ChemiDoc MP Imaging System with Image Lab
Software, PC or Mac, includes darkroom,
UV transilluminator, epi-white illumination, camera,
power supply, cables, Image Lab Software
Accessories
170-8283
Red LED Molecular Kit, pkg of 2 epi-red LED
modules and 1 red emission filter, for use with
applications requiring red fluorophore detection
170-8284
Green LED Molecular Kit, pkg of 2 epi-green LED
modules and 1 green emission filter, for use with
applications requiring green fluorophore detection
170-8285
Blue LED Molecular Kit, pkg of 2 epi-blue LED
modules and 1 blue emission filter, for use with
applications requiring blue fluorophore detection
170-8289
White Light Conversion Screen, for viewing
Coomassie, silver stain, and other colorimetric
gel stains
170-8182
XcitaBlue Conversion Screen, includes view
goggles; blue conversion screen for viewing
SYBR® Green, SYBR® Safe, GFP, Flamingo, and
other fluorescent gel stains
170-8183
XcitaBlue Conversion Screen and Filter, includes
view goggles and SYBR® Safe Filter (170-8075,
560DF50); blue conversion screen for viewing
SYBR® Green, SYBR® Safe, and other fluorescent
gel stains
170-8083
Filter 520DF30 62 mm, for SYBR® Green/GFP/SYBR®
Gold/fluorescein
170-8098
254 nm UV Lamps, pkg of 6
170-6887
365 nm UV Lamps, pkg of 6
170-8097
Standard 302 nm UV Lamps, pkg of 6
170-8089
Mitsubishi Thermal Printer
170-7581
Mitsubishi Thermal Printer Paper, 4 rolls
170-8184
Gel Alignment Templates, pkg of 3
Detector
Supercooled CCD
Image resolution
4 megapixels
Software
170-9690**
Image Lab Software, PC or Mac, for automated
image capture, optimization, and 1-D data analysis
Pixel size (H x V)
6.45 x 6.45 µm
** Included with the imaging system.
Cooling system
Peltier
Camera cooling temperature
–30°C absolute and regulated
Filter holder6 positions (5 for filters, 1 without filter
for chemiluminescence)
Emission filters
1 included (standard), 4 optional
Dynamic range
>4.0 orders of magnitude
Pixel density (gray levels)
65,535
Instrument size (L x W x H)
36 x 60 x 96 cm
Instrument weight 32 kg
Operating Ranges
Operating voltage
110/115/230 VAC nominal
Operating temperature
10–28°C (21°C recommended)
Operating humidity
<70% noncondensing
* U.S. patent 5,951,838.
Scan this QR code to learn more about
the ChemiDoc MP Imaging System, or visit
biorad.com/chemidocmp.
Alexa Fluor, Coomassie Fluor Orange, Pro-Q, Qdot, SYBR, and SYPRO are
trademarks of Life Technologies Corporation. Coomassie is a trademark
of BASF Aktiengesellschaft. Cy is a trademark of GE Healthcare Group
Companies. DyLight and Krypton are trademarks of Thermo Fisher Scientific.
GelRed is a trademark of Biotium, Inc. IRDye is a trademark of LI-COR
Biosciences. Mac is a trademark of Apple Inc. Mitsubishi is a trademark of
Mitsubishi Companies. Q-Plex is a trademark of Quansys Biosciences.
Bio-Rad Laboratories, Inc. is licensed by Life Technologies
Corporation to sell SYPRO products for research use only,
under U.S. patent 5,616,502.
Bio-Rad
Laboratories, Inc.
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Life Science
Group
Bulletin 6133 Rev D
US/EG
14-2254
0315
Sig 1214
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