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NH DyeAGNOSTICS is partnering the Scientific Meeting
N-term 2017: Proteostasis via the N-terminus
September 11-13th 2017
located at the Leibniz-Institute of Plant Biochemistry in Halle, Germany.
More meeting details…
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List of speakers
Christopher Brower, Nico Dissmeyer, Mark Ditzel, Jürgen Dogmen, David Dougan, Ruth Geiss-Friedlander, Kris Gevaert, Daniel Gibbs, Carmela Giglione, Emmanuelle Grafite, Richard Fahlman, Emily Flashman, Michael Holdsworth, Yong Tae Kwon, Dominique Leboeuf, Francesco Licausi, Thierry Meinnel, Angelika Mustroph, Jos Schippers, Hyun Kyu Song, Takafumi Tasaki, Frederica Theodoulou, Klaas van Wijk, Rens Voesenek, Markus Wirtz, Martin Zenker
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The group of the organizer, Dr. Nico Dissmeyer has successfully used the patented SPL technology for quantitative Western Blot analysis (Faden et al. 2016*).
Abstract of our meeting poster:
Real-time Normalization of Western Blots: Smart Protein Layers
Jana Heise1,2, Jan Heise1
1 NH DyeAGNOSTICS GmbH, Weinbergweg, 23 06120 Halle, Germany
2 Weinberg Laboratories, Weinbergweg 23, 06120 Halle, Germany
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Over decades, Western blotting is a method to detect a target protein in a complex sample. The technique includes protein separation, transfer to a membrane and several washing and antibody incubation steps.
Reliable comparison of detected target signal intensities between different samples is still a problem. It requires an appropriate way of normalization to assure that the reported fold changes of the target protein are not an artifact of a reference signal and/ or experimental errors.
Smart Protein Layers (SPL) is a add-on kit for the detection of sample proteins present on the blot at the same time the target is immuodetected. It is based on two components: i) fluorescent SPL Labels, bound to the sample protein prior separation, for the visualization of all proteins present on the blot at this time ii) bi-fluorescent SPL Standards to monitor labeling efficiency and to provide additional information about the sample and to enable automated data evaluation.
With SPL the determined expression of a specific target protein can be normalized to its corresponding sample, providing a reliable and precise tool for the comparison of fold changes between different samples.
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*Faden F., Eschen-Lippold L., Dismeyer N..2016. Normalized Quantitative Western Blotting Based on Standardized Fluorescent Labeling. Methods Mol Biol. 1450:247-58.
