High mobility group box 1 promotes tumor cell migration. With these systems, most of the work falls into planning the experiments to get the most out of each run and interpreting the data, she adds. Whatever your imaging application the sapphire biomolecular imager and the powerful sapphire capture software will deliver high quality quantifiable data. The odyssey twocolor protein molecular weight marker is prestained and fluoresces in the 700 and 800 nm channels of the odyssey system.
Scan on odyssey using protein gel preset in odyssey software and scan in 700. The compact optical system uses freespace lasers to concentrate intensity at the flow cell. Image studio software work area folder best practices canning a mouse on. Being able to double label with the 700 and 800 ir laser also allows us to save blots and time. The marker was designed for use with polyacrylamide gels and can be used to visualize the progress of protein separation during electrophoresis, to monitor transfer efficiency to western blot membrane, and to. The licor platform has broad applicability for quantitative western blots, protein gel imaging, and incell westerns, providing for quantitative analysis and a wide linear dynamic range that is not available with traditional chemiluminescent methods. Incell western assays to evaluate hantaan virus replication. The odyssey clx combines precision laser illumination, large scan area, and high image resolution to deliver accurate, replicable data capture. Incell western assay, in cell elisa, analyzes target proteins in fixed cells in plates using licor secondary antibodies and infrared imaging on odyssey. Imaging software is then used to compare the signal generated by the bands detected on the western blot. Licor, odyssey, mpx, incell western, and irdye trademarks contained in the software product are trademarks or registered trademarks of licor, inc. Direct infrared detection is available on two different systems from licor biosciences the odyssey infrared imaging system and the aerius automated infrared imaging system. Odyssey clx infrared imaging system operators manual. This leads to better data that you can be confident in.
Immunoblots, total cell lysates were immunoblotted for the indicated endogenous proteins. Quantitative immunofluorescent assays using infrared. Bd facsuite software delivers a seamless workflow from system setup, through data acquisition, analysis, and shutdown. Licor, odyssey, mpx, incell western, and irdye trademarks contained in the software product are trade marks or registered trademarks of licor, inc. Molecular imaging on the licor odyssey imaging system.
Nearinfrared fluorescent western blot, incell western assay, emsa, protein gel documentation, nucleic acid gel documentation, tissue section imaging, ingel westerns, rnai analysis. Gels and blots can be detected for 30 seconds, 2 minutes or 10 minutes. Variability in the manual processes that used to impact reproducibility, quantitation, time to result and overall data reliability is eliminated. Odyssey clx infrared imaging system from licor biosciences. The incell western icw assay is a quantitative immunofluorescence assay performed in microplates optimized for 96 or 384well format that combines the specificity of western blotting with the replicability and throughput of elisa. Odyssey clx imaging system minimize variation and maximize precision with highsensitivity nir fluorescent detection optics engineered to enable definitive analysis of protein expression changes. Qwb data was analysed using odyssey software as per manufactures guidelines and as previously described. After weeks of selection, cells were analyzed by western blot for hmgb1 and perk levels using licor odyssey software. Software upgrades are free stay current for the latest innovations. Odyssey clx manuals, application notes, software, and webinars. All is not quiet on the western front the scientist. Licor biosciences, odyssey infrared imaging system b446.
Data was graphed and statistical comparisons carried out using graphpad prizm as previously described. Fluorescent western scanning imaging system licor odyssey c l x infrared imaging system. For the complete list of video tutorials and support. One major advantage of licor odyssey system over normal systems is that you can use at least 2 primary antibodies e. The licor odyssey fc imaging system is easy to use and the operation is selfexplanatory.
The sapphire biomolecular imager quantitative western. An on cell western ocw assay is a cell based assay that enables quantitative monitoring of cell surface protein expression. Get an image acquisition software designed for accurate western blot. With odyssey clx, you can get consistent, reproducible digital images, without the hassles and unpredictability of film. Nearinfrared fluorescent western blot, in cell western assay, emsa, protein gel documentation, nucleic acid gel documentation, tissue section imaging, in gel westerns, rnai analysis.
Molecular imaging on the licor odyssey imaging system mousepod. Licor, incell western, mpx, newblot, odyssey, and irdye are registered. Odyssey clx application and technical notes show the versatility of the clx. Licor odyssey gel imaging scanner university of washington. Licor last week introduced the aerius automated infrared imaging system for cellbased protein discovery using infrared platebased assays according to the company, the aerius builds upon the odyssey twochannel ir detection system, adding a. Page 2 how to use the image studio software incell western analysis. Used other licor odyssey imager for sale dotmed listing. Licor image studio software for accurate data quantification. Automatically finds wells for 96 and 384well microplates, and performs ratiometric calculations using the two imaging channels of the odyssey system. The new odyssey clx infra red imaging system from licor biosciences improves your quantitative analysis for detecting proteins on membranes, in cells, in tissue, invivo etc. Which is the best and more sensitive western blot imaging. Jess gives you four different ways to analyze proteins. Analyze incell western assays with licor image studio software. The 640089 kit contains a capped cylinder 3 cm dia.
Licor s aerius automated infrared imaging system, odyssey. Image studio lite is a free software package from licor biosciences aimed at life scientists that want to analyze gels, western blots, dot blots, and other similar lab outputs. The licor odyssey 9120 delivers accurate and reproducible results with western blot and more. The icw assay is a cellbased technique for intracellular protein detection that is characterized by high rapidity, accuracy, sensitivity, and reproducibility egorina et al. These tools include the ability to perform cell count normalization, percent response, and zfactor analysis. Odyssey applications quantitative western blots coomassie stained gels cell. Both systems are designed for incell westerns and other membrane and platebased assays. Studies using confocal microscopy have previously shown that cb1 internalizes following exposure to cb1speci.
Pathway analysis with the in cell western assay learn more about in cell westerns. Although the lite version is free, there is a more comprehensive paid version of the software that aims to easily integrate with the apparatus that licor also sells. Membranes were scanned on the licor odyssey ir imaging instrument, and densitometric analysis was executed via the imagej software. The major advantage with this solution is accurate quantification over a. Quantitative immunofluorescent assays using infrared fluorescence. Odyssey clx imager page 3 placement in the laboratory the odyssey clx infrared imaging system weighs approximately 33 kg 72 lbs. Licor odyssey sa infrared imaging system biosurplus. Mar 04, 2014 this video tutorial demonstrates the image studio 4. Be sure you are meeting the new stringent publishing and granting agency requirements. Try empiria studio software today and enhance your odyssey workflow with the only western blot analysis software.
For quantitation, entire lanes in the blot were selected and fluorescence in each channel quantitated using the licor software. The consistent digital images produced by the imaging system cut out the hassle of film, while providing clear near infrared results. Control lanes included extracts from cells treated without puromycin. Information on odyssey software can be found both in this manual and in the odyssey user guide. The licor odyssey western blot delivers quantitative accuracy for target protein detection by using the platforms infrared technology to match or exceed chemiluminescent detection sensitivity. Automated western blot infrared imaging system model 9260.
The major advantage with this solution is accurate quantification over a broad dynamic range, up to 6 orders with high sensitivity. Licor odyssey platform for quantitative western blots. The kits feature irdye 800 cw and irdye 680 labeled secondary antibodies, odyssey blocking buffer, and pvdf membranes. The licor odyssey imaging system offers in vitro and in vivo imaging with its proprietary nearinfrared imaging system. Licor odyssey clx college of education and human sciences. Blots were scanned using the licor odyssey scanner licor biosciences, lincoln, ne and each channel visualized independently or simultaneously. Pathway analysis with the incell western assay licor. To narrow this gap, in cell western icw assays have been applied to monitor hantavirus replication kinetics and assess viral titers. Which is the best and more sensitive western blot imaging system. Western blot detection methods this protocol is designed to help you achieve success with nir western blot detection methods.
Used licor odyssey sa infrared imaging system 3014309. The licor website provides updates on new developments in infrared imaging and. We continue to provide chambers, light sources, service, and support for li6400xts. Imaging is rapid, with adjustable signal intensity, and integration.
It has two lasers, which operate at 680 and 780nm, for the detection of infrared fluorescent dyes. The in cell western icw assay is a quantitative immunofluorescence assay performed in microplates optimized for 96 or 384well format that combines the specificity of western blotting with the replicability and throughput of elisa. Typical workflow, basic considerations for success, and sample protocol for. By using digital images, strong and weak bands are detected on the same blot, saving labs time by not waiting on film to develop or the uncertainty whether a weak band requires multiple exposures to confirm. The odyssey clx is the next generation multifunctional imaging platform that can provide a wide range of applications. Free western blot quantification software licor biosciences. All measurements were normalized to the dmso0 h sample with relative value of 1. In cell western assay cell fixationpermeabilization.
Detect and quantify target proteins localized to the cell surface. The li6800 includes easytolearn software that makes it easier to understand the complexities of photosynthesis measurements. Best of all, the transition is simplethe same antibodies you use for western blotting protocols can be used with simple western assays. Perform many different applications, including nearinfrared fluorescent western blots, cellbased assays such as incell western or oncell western assays, protein and nucleic acid gel imaging, tissue section imaging, fluorescent emsas, and more. Analyze your data right away and plan your next experiment. It can analyze multiple samples quickly and quantitatively, and it eliminates the use of radioactivity for the assay. Western blots, incell westerns, coomassie stained gels, dna gels,fluorescent gelshift assays, tissue imaging, in vivo imaging, whole organ imaging, protein arrays, and more serial dilutions 10 ng to in the licor lot. Odyssey clx application protocols manual purdue biological. As with any western blot method, the quality of your antibody can be a limiting. The auction is held in our seminar room during break and lunch. Quantitative western blots, protein gel imaging, and incell westerns.
It gives a basic overview of the functionality and i run through the analysis of a gel. Odyssey clx molecular imaging workstation from licor. A good place to start is to read the overview of odyssey in this chapter. Compare odyssey clx imaging system from licor biosciences on. Pathway analysis with the incell western assay learn more about incell westerns. The icw assay is a cell based technique for intracellular protein detection that is characterized by high rapidity, accuracy, sensitivity, and reproducibility egorina et al. An oncell western ocw assay is a cellbased assay that enables quantitative monitoring of cell surface protein expression.
The new odyssey clx infra red imaging system from licor biosciences improves your quantitative analysis for detecting proteins on membranes, in cells, in tissue, in vivo etc. Odyssey clx application protocols manual vanderbilt university. Licor odyssey clx the odyssey clx is the next generation multifunctional imaging platform that can provide a wide range of applications. The licor website provides updates on new developments in infrared imaging and research results using the odyssey and aerius systems. Since the odyssey is a two channel instrument it can record data for either wavelength separately or both wavelengths simultaneously. The licor odyssey 9120 imaging system gives your lab clear sharp digital results without the background fluorescence. However, the sapphire biomolecular imager is a next generation laser scanning system which offers the best of both worlds. Imager biorad v3 western blot university instrumentation. Graphs, transcript levels as measured by sybrgreen qrtpcr and protein levels as quantified by licorodyssey software on immunoblots are compared for the indicated proteins.
Odyssey software accessory packages incell western analysis software for in vitroassays in microplate format. Licor biosciences introduces the odysseyr twocolor nir. Development and validation of an incell western for. Oct 21, 2016 blots were scanned using the licor odyssey scanner licor biosciences, lincoln, ne and each channel visualized independently or simultaneously. When initial screen has loaded, click on file and select new from the dropdown menu. Licor image studio lite free densitometry software youtube. The sapphire biomolecular imager quantitative western blots. A microplatebased nonradioactive protein synthesis assay. The bd facsverse flow cytometer is designed to support cell analysis for research applications using up to 10 parameters. By using nir fluorescence detection, this imaging system greatly simplifies your western blot process by eliminating the need to incubate your substrate or wait for film. A complete list of incell western assay protocols oneblot western. This video tutorial demonstrates the image studio 4. The instrument should be placed on a laboratory bench that is sufficiently sturdy to bear its weight. This is a quick look at the free densitometry software from licor.
The licor odyssey allows us to veiw our western blots quickly and efficiently. Facilities and equipment howard university department of. Li6400xt portable photosynthesis system licor environmental. For western blotting, 3 levels of sensitivity have been optimized. Jess chemiluminescent and fluorescent western blotting. In addition to twocolor infrared and in gel quantitative western blots, bioagilytix uses our licor odyssey platform for cell based assays, protein detection, nucleic acid detection, protein arrays, rnai analysis, and more.
Total protein analysis as a reliable loading control for. Densitometers and analysis software in order to make a permanent record, or to get an objective measure of the signal generated on a western blot, a densitometer is used to scan the blot or film. All blots were treated equally, with the exception of blocking reagent. Output data are easily managed tiff files, with no need for systemspecific software. Mar 18, 2015 this is a quick look at the free densitometry software from licor.
High sensitivity, with quantitation, is stateoftheart fold over licor imaging. See some published examples of in cell western assays. Licor last week introduced the aerius automated infrared imaging system for cell based protein discovery using infrared platebased assays according to the company, the aerius builds upon the odyssey twochannel ir detection system, adding a microplate reader capable of handling up to 30 in a single run. If you have the need to do chemiluminescent western blots and quantitative infrared westerns plus other applications such as platebased assays or small animal imaging, this package is just the right fit. The software can take a bit longer to learn, depending on how complex the experiment is, liliental says. The licor odyssey clx imaging system gives your lab clear sharp digital results in as little as 6 minutes. The odyssey clx molecular imaging workstation includes odyssey clx infrared imaging system with unlimited image studio software user licenses pn 914003 and the pearlimpulse small animal imaging system 940003 with unlimited user software licenses. In addition, software functions related to the odyssey mousepod accessory are discussed in the odyssey in vivo imaging guide. Get consistent, reproducible digital images, without the hassles and unpredictability of film. Leveraging this platforms capabilities, bioagilytix is able to perform accurate, reproducible, quantitative western blot assays. Two different wavelengths for western blot fluorescent dyes 680 and 800nm and pcr gels dyed with gel red detection 600 nm are available. Read the entire protocol carefully before beginning your optimization experiments. The odyssey infrared imaging system allows multiplex western blotting and accurate protein detection and quantification using secondary antibodies labeled with irdye nearinfrared nir fluorescent dyes. Applicationsnearinfrared fluorescent western blot, incell western assay.