With high precision and sensitivity, the Odyssey M Imager is all encompassing for even the most advanced research initiatives. The Odyssey M Imager is the most powerful imager currently offered for fluorescent (i.e., near-infrared (NIR) and visible), luminescent, and white light imaging. This type of multicolor analysis not only reduces workflow significantly by minimizing the need for multiple blots, but also makes the quantitation of multiple proteins much more reliable.Seek Discovery and Leave Limitations Behind with M In this example, multiplex detection facilitates the normalization of fluorescent signals and allows us to distinguish proteins that co-migrate in a single gel lane, enabling the identification and quantitation of posttranslational modifications when specific primary antibodies for these modifications are available. Using protein-specific primary antibodies and Qdot® secondary antibody conjugates, we have probed a single western blot for two proteins of interest (EGFR and phosphorylated EFGR), as well as a third protein (GAPDH) for normalizing signals (Figure 2). More advanced fluorescence imagers, such as the Fujifilm® LAS-4000 gel imager, enable higher-order multiplexing and quantitation. Qdot® fluorescence can be detected on many commercially available fluorescence imagers, including relatively inexpensive gel imagers equipped with a UV light box and appropriate emission filters-e.g., the E-Gel® Imager with the Qdot® 625 filter. Qdot® nanocrystals are exceptionally bright and photostable fluorophores that can be excited at any wavelength below their emission (though most efficiently by UV, violet, or deep-blue light). WesternDot™ 800 anti–mouse IgG ( W10815) and anti–rabbit IgG antibody ( W10816) conjugates.WesternDot™ 625 anti–mouse IgG (W10805, W10808), anti–rabbit (W10806, W10809), and anti–goat IgG (W10807) antibody conjugates (available by phone order only until December 15, when they will also be available online).We offer the following WesternDot™ secondary antibody conjugates: Additionally, these WesternDot™ conjugates have reduced intensity differences between the different colors, simplifying multiplex applications. Furthermore, our new WesternDot™ antibodies utilize Qdot® nanocrystals enhanced with VIVID® technology, making them brighter than the original Qdot® reagents. Qdot® secondary antibody conjugates provide a powerful alternative to traditional methods for western blot detection. Multiplex westerns with Qdot® antibody conjugates Moreover, data analysis is greatly simplified because you can directly compare signals from protein bands that have been exposed to identical electrophoresis and transfer conditions. A multicolor western blot saves time and sample by eliminating the need to produce duplicate gels and blots or to strip and reprocess a single blot with a different antibody probe. Tremendous flexibility in experimental design is afforded by the availability of several different long-wavelength Qdot® and Alexa Fluor® fluorophores, facilitating the detection of multiple proteins on a single western blot (Figures 2 and 3). Importantly, probes with long-wavelength (red to near-IR) emission allow signal collection beyond the typical autofluorescence emitted by standard nitrocellulose and PVDF membranes, resulting in low background fluorescence and superior signal-to-noise ratios. When paired with a fluorescence imager, these detection reagents provide sensitive and quantitative analysis of proteins blotted or spotted onto membranes. Qdot® and Alexa Fluor® secondary antibody conjugates are revolutionizing western blot detection. Advantages of fluorescent probes for western blots
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