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Multiplexing LI-COR® IRDye® 680LT and IRDye® 800CW Using the G:BOX Chemi and PXi Imaging Systems from Syngene

Syngene
29 Apr 2015

This application note demonstrates that users can successfully multiplex between LI-COR dyes IRDye 680LT and IRDye 800CW using Syngene’s G:BOX Chemi and PXi imaging systems. One of the main advantages of using a G:BOX Chemi and PXi system is the faster imaging exposure times compared to that of the Odyssey Infrared Imager and also that the G:BOX Chemi and PXi systems offer great versatility enabling the imaging system to be used for a variety of different applications.

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G:BOX Chemi XT4

Syngene

For high performance imaging, the new generation of G:BOX Chemi gel doc systems provide the user with an extended range of applications covering fluorescence and chemiluminescence. All these systems now utilise Syngene’s groundbreaking GeneSys automatic control software. Driven from a database containing hundreds of capture protocols, full manual control is available should you wish to use it. The G:BOX Chemi series features cameras with resolutions of 4m, 6m or 9m pixels.  All have cooling which reduces the small amount of electronically generated noise to produce outstanding images with minimal background interference. These new generation cameras also have higher quantum efficiencies (QE) than most, helping the system to achieve outstanding levels of sensitivity. Whether using the system for standard fluorescence applications such as DNA, or for chemiluminescence imaging, the G:BOX Chemi will give you high quality results. In addition, by using the optional LED modules users can work with a unique automated multiplexed function capable of using up to 5 channels. Other LED modules can be used to turn the G:BOX Chemi into a highly sensitive IR capture system.  An edge lighting option can also be used for 2D gel capture including the use of DIGE gels. The stylish design with modular construction includes motor-driven lenses, filter wheels with the option of including a range of lighting choices for both Epi and transillumination applications. Syngene has just introduced the new high intensity HI-LEDs option for the G:BOX range, resulting in superior imaging performance for your fluorescence applications. Check the status of an image capture and access your recently captured images from any computer, smartphone or tablet on the same network as G:BOX Chemi XT4 gel doc system using the new "Status Link" feature. This is ideal for any busy laboratory. The system comes complete with unlimited copies of GeneTools analysis software.

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ElectrophoresisElectrophoresis equipment is used to separate mixtures of protein, DNA or RNA, based on their electric charge, size and other physical characteristics, by passing them through a medium such as a polyacrylamide gel, an agarose gel or a capillary tube. Electrophoresis equipment includes horizontal and vertical gel electrophoresis chambers, isoelectric focusing systems, 2D electrophoresis and capillary electrophoresis instruments. Precast gels with a gradient can be used or gels can be hand cast. Samples are run alongside ladders or markers to identify the approximate size of a molecule. Separated proteins and nucleic acids in the gel can be stained or probed with fluorescent markers and then imaged and detected using gel documentation instruments, transilluminators, densitometers and scanners. Find the best electrophoresis equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Western BlottingWestern blotting equipment is used to transfer and identify specific proteins within a sample, reveal protein modifications, as well as give a semi-quantitative estimation of their concentration. Western blotting equipment includes all apparatus necessary to transfer proteins from gel to membrane and subsequent processing steps. Protein transfer can be performed by electroblotting with wet, semi-dry and dry transfer systems onto nitrocellulose and PVDF membranes. Blocking, washing and labeling of membranes follows, involving buffers, blocking reagents, blotting / incubation trays, labeling reagents, immunoblotting assays, antibodies and conjugates. Automated equipment for these steps is available to accelerate your lab workflow. Finally, detection and imaging of proteins can be conducted using gel documentation and imaging systems. Find the best western blotting equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.Protein PurificationProtein purification is a vital step in drug discovery, therapeutics, biotech and life science research. The purification process typically involves subcellular or membrane protein extraction with cell lysis kits, separation of proteins from cell debris by filtration or spin columns, and the isolation of proteins of interest from other proteins and impurities with affinity purification (including fusion protein tags and antibody binding proteins A, G and L), immunoprecipitation or chromatographic methods, such as ion exchange, size exclusion and immobilized metal affinity chromatography. All purification methods come in multiple formats for your laboratory needs, including agarose or magnetic beads, resins, columns and filter plates. Find the best protein purification equipment in our peer-reviewed product directory: compare products, check customer reviews and receive pricing direct from manufacturers.FluorescenceThe emission of fluorescence occurs when a photon of energy is supplied to a fluorescent chemical compound by an external source, causing it to become excited. Fluorescence can be detected and measured for different purposes using microplate readers, fluorescence microscopes, fluorescence scanners, and flow cytometers.Gel ImagingGel imaging captures high-resolution images of electrophoresis gels to analyze DNA, RNA, and proteins. Essential in molecular biology, this technique helps visualize results for accurate quantification and documentation. Discover top imaging systems and compare user-reviewed options in our directory.MultiplexingMultiplexing refers to the ability to measure multiple targets or analytes simultaneously in a single experiment. This technique is valuable for high-throughput screening, diagnostics, and complex assays, as it increases efficiency and data quality. Browse our peer-reviewed product directory to find the best multiplexing products, compare tools, check reviews, and get pricing directly from manufacturers.

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