Accurate, Reproducible Western Blots with the Odyssey CLx

Odyssey® CLx
Imaging System

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Get clearer answers with accurate, reproducible Western blots and more

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Do More with Digital Fluorescence

Get consistent, reproducible digital images, without the hassles and unpredictability of film – so you can analyze your data right away and plan your next experiment.

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See More with Clear Western Blot Images

See both strong and faint bands clearly in the same image, with great sensitivity and no image saturation. Multiplex with two fluorescent colors to see even more data.

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Discover More with Richer, More Complete Data

Capture all the detail and complexity of your data. With 6 logs of linear dynamic range, your results are more reproducible. Near-infrared (NIR) fluorescence delivers consistent signals that aren’t affected by timing.

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Do more with your time

Skip the mess and inconsistency of film, substrates, and the darkroom. Direct near-infrared fluorescence gives you high-quality images without the subjectivity or hassles of multiple film exposures.

 

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Simplify your Western blot protocol. Save time and minimize the number of manual steps that can introduce variability. NIR fluorescence detection doesn’t require chemiluminescent substrates or film. Just place your blot inside the Odyssey CLx imager and start collecting data in digital format.

Do more with your funding

Film, chemiluminescent substrates, and darkroom expenses eat up resources that you could be using to move your research forward. Film processing chemicals and rinse waters are hazardous wastes that require special disposal. And a single X-ray film processor consumes an average of 788,000 gallons of water per year.1, 2

“The equipment is versatile and easy to use. It saved my group a lot of money as compared to traditional Western blotting.”

Clifford Nwaeburu, German Cancer Research Center, SelectScience® Review

Save thousands in ongoing costs with the Odyssey CLx.
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Save thousands in ongoing costs with the Odyssey CLx.
Save thousands in ongoing costs with the Odyssey CLx.

Save thousands in ongoing costs with the Odyssey CLx.
Digital imaging eliminates typical darkroom expenses like processing chemicals, developer maintenance, and film.

 

Image and analyze your
samples quickly

Analyze your data right away – it’s already in digital format. One digital file contains all your image data. Record-keeping is easy and multiple exposures are a thing of the past.

To image and analyze many samples efficiently, use the Odyssey CLx to scan up to 9 mini-blots, 6 microplates, or 30 slides at the same time.

Go where your experiments take you

Cell-based assays, protein arrays, gel shift assays, tissue section imaging, and more are at your fingertips with near-infrared fluorescence.

“We are finding that we can use the Odyssey CLx for just about everything protein-related we do in the lab.”

Dr. Jeremy Chambers, Florida International University

In-Cell Western Assay

In-Cell Western™ Assay

NIR Fluorescent Western Blot

NIR Fluorescent Western Blot

EMSA / Gel Shift Assay

EMSA / Gel Shift Assay

Tissue Section

Tissue Section

Image courtesy of C. Kearn, University of Washington

See More with Clear Western Blot Images

Get the full story in a single image

One digital image file contains all your data. See both strong and faint bands clearly in a single image, without image saturation, “blowout”, or sacrificing sensitivity.

Increase the consistency and reproducibility of your results by capturing a single image, rather than comparing multiple exposures captured under variable conditions.

“The superior dynamic range and sensitivity allows me to confidently report my data.”

Lars Engstrom, Mirati Therapeutics, SelectScience® Review

Odyssey Clx Western Blot Image Sequence 1 Odyssey Clx Western Blot Image Sequence 2 Odyssey Clx Western Blot Image Sequence 3 Odyssey Clx Western Blot Image Sequence 4 Odyssey Clx Western Blot Image Sequence 5 Odyssey Clx Western Blot Image Sequence 6 Odyssey Clx Western Blot Image Sequence 7 Odyssey Clx Western Blot Image Sequence 8 Odyssey Clx Western Blot Image Sequence 9 Odyssey Clx Western Blot Image Sequence 10 Odyssey Clx Western Blot Image Sequence 11 Odyssey Clx Western Blot Image Sequence 12 Odyssey Clx Western Blot Image Sequence 13 Odyssey Clx Western Blot Image Sequence 14 Odyssey Clx Western Blot Image Sequence 15 Odyssey Clx Western Blot Image Sequence 16 Odyssey Clx Western Blot Image Sequence 17 Odyssey Clx Western Blot Image Sequence 18 Odyssey Clx Western Blot Image Sequence 19

Show different views by adjusting the image display

One image file contains the full range of your data. Choose the best image display for further analysis, based on your protein of interest. Changing the image display settings never alters your raw data or signal intensities.

Get better sensitivity with low image background

High signal-to-noise ratios give you more confidence in your data, and can help you reliably detect subtle changes between samples. Low background fluorescence at near-infrared (NIR) wavelengths means high sensitivity for your blots. In the visible spectrum, detection sensitivity is limited by high autofluorescence of blotting membranes, plastics, and biological materials.

Powerful, precise laser excitation and specialized optics are the keys to high signal-to-noise ratios and outstanding image quality. Other imagers use LEDs or diffuse white light sources that provide weak, non-specific excitation light and limit sensitivity.

Visible region (Vy3) High autofluorescence Neasr-infrared region (700nm) Low autofluorescence

NIR fluorescence imaging gives you low membrane fluorescence and high sensitivity. Visible fluorescent dyes like Cy3 have poor sensitivity, because membrane autofluorescence is so strong. Blots show detection of phospho-ERK with NIR fluorescence (700 nm) and with ECL Plex reagents.

See your bands reliably with stable fluorescent signals

Near-infrared fluorescent signals are stable and reproducible over time. Timing and enzyme kinetics don’t affect your results, so you can image your blots when it’s convenient for you. You can even re-image the same blot later and see the same results.

Fluorescent signals are stable and unaffected by timing, so you can compare band intensities with confidence.
Fluorescent signals are stable and unaffected by timing, so you can compare band intensities with confidence. NIR fluorescent signals are stable for months, because no enzymes or substrates are used. Direct detection is performed using secondary antibodies labeled with IRDye near-infrared fluorescent dyes, or other commercially-available dyes. You’ll typically lose signal in less than an hour with ECL substrates and indirect enzymatic detection.

See multiple targets on
the same blot

With multiplex fluorescence, you can detect two protein targets in each sample lane, with great sensitivity in both fluorescence channels. Consistency and reproducibility are improved because you can account for lane-to-lane variation in loading and transfer without stripping and re-probing.

“The infrared imaging system allows detection of multiple proteins on a single blot, without the need for stripping and re-probing.”

Bond et al (2008) Biol Proced Online. 10: 20–28.

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700nm
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800nm
700nm and 800nm Western Blot

Multiplex to detect two different protein targets in each sample lane. Use secondary antibodies labeled with spectrally-distinct NIR fluorescent dyes to get more data from your blot. View, adjust, and analyze your results as a merged image, or as separate 700 nm and 800 nm channel images in pseudo-color or grayscale.

See your strong bands accurately

Digital fluorescence shows your strong signals as they really are. Film and CCD imagers only show you part of the picture, because data are lost when images become saturated. To see the whole picture, you need enough capacity (dynamic range) to consistently document your strongest bands.

“When saturated, film exposures can also hide sample-to-sample variations in high-abundance proteins.”

Janes (2015) Science Signaling 8(371): rs2

 

Get deeper data capacity with digital fluorescence. The Odyssey CLx Imager gives you a stunningly wide linear dynamic range – in other words, a very deep data capacity. Film has a shallow data capacity and cannot accurately record strong signals. Once that capacity is reached, all additional signal (read: your data) is simply lost. Many CCD imagers are also limited by data capacity and image saturation.

Discover More with Richer,
More Complete Data

Detect and analyze all of your bands, faint and strong

You ask big questions, and your detection method should be able to handle all the answers. Now you can capture all of your data in a single image. Strong and faint bands are accurately documented in just one scan by the Odyssey CLx Imager.

With an unprecedented 6 logs of linear dynamic range, you’ll never lose data because of image saturation. Other detection methods can’t handle the challenge and let some of your data slip away.

“The ability to quantitate those bands is just amazing!”

Dr. Wendy L. Picking, Associate Professor, Oklahoma State University

Capture and analyze all of your data, without image saturation. Capture and analyze all of your data, without image saturation. Capture and analyze all of your data, without image saturation. Capture and analyze all of your data, without image saturation. Capture and analyze all of your data, without image saturation. Capture and analyze all of your data, without image saturation. Capture and analyze all of your data, without image saturation.

Capture and analyze all of your data, without image saturation. When your detection method has the capacity to collect all of your data, you can detect the differences between strong bands. Digital NIR fluorescence reveals the complete picture. With film you can’t tell when saturation begins, and it’s different in every exposure. Strong bands are all similarly dark, and densitometry can’t show you the differences. When a CCD imager reaches saturation (blue pixels), the results are inaccurate and not suitable for analysis (dotted line).

Find out what’s really happening in your samples

Your Western blot data should reflect real changes in your samples, not the timing of your detection. Stable near-infrared fluorescent signals give you consistent, reproducible answers – no matter when you image.

But chemiluminescent detection and other enzyme/substrate methods are “moving targets”. Signals are dynamic and constantly changing, with inconsistent enzyme kinetics across the blot. Data output is heavily dependent on timing, so relative comparisons of band intensity may give very different results for different time points. The answer you get depends on when you ask.

With NIR fluorescence imaging, you can be more confident that the changes you see on your Western blots reflect actual changes in the biochemistry and composition of your samples.

“Just because we can put numbers on an image does not imply that we should.”

Janes (2015) Science Signaling 8(371): rs2

Cumulative band intensity

Timepoint 1 Timepoint 2 Timepoint 3

Stable fluorescent signals deliver consistent, proportional results for quantitative analysis. Relative comparison of band intensity is based on consistent, reproducible signal output. With NIR fluorescence, relative band intensity is constant and proportional across time, every time. But with chemiluminescent detection, the enzymatic reaction is constantly changing. You’ll see different band intensities at different times and get different answers. These time-dependent changes reflect the limitations of chemiluminescence, not real changes in your samples.

See the Odyssey CLx in action

Do more with consistent, reproducible digital imaging and the flexibility to perform a variety of assays and experiments. See the benefits of stable NIR fluorescence, and detect multiple targets in the same lane. Get the most accurate results with over 6 logs of linear dynamic range, so you can make more discoveries.

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“The brilliant signal-to-noise ratio in combination with the ability to truly quantify the data is really outstanding.”

Geir Bjørkøy, University College of Sør-Trøndelag

How It Works

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  1. US Environmental Protection Agency. Laboratories for the 21st century: best practices. Office of Administration and Resources Management. DOE/GO-102005-2008 (2005).
  2. Cohen, R et al. Making every drop work: increasing water efficiency in California’s commercial, industrial, and institutional sector. Natural Resources Defense Council (NRDC) Issue Paper (2009).
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