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<h1 id="firstHeading" class="firstHeading mw-first-heading"><span class="mw-page-title-main">SYPRO Orange</span></h1>
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<p><b>SYPRO Orange</b> ist ein <a href="Fluoreszenzfarbstoff" class="mw-redirect" title="Fluoreszenzfarbstoff">Fluoreszenzfarbstoff</a> vom <a href="Cyanine" title="Cyanine">Merocyanin</a>-Typ,<sup id="cite_ref-Demchenko_1-1" class="reference"><a href="#cite_note-Demchenko-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup> der im Zuge einer <a href="Proteincharakterisierung" title="Proteincharakterisierung">Proteincharakterisierung</a> zur <a href="Proteinf%C3%A4rbung" title="Proteinfärbung">Proteinfärbung</a> verwendet wird,<sup id="cite_ref-EP0774122_2-1" class="reference"><a href="#cite_note-EP0774122-2"><span class="cite-bracket">[</span>2<span class="cite-bracket">]</span></a></sup> meistens in einer <a href="SDS-PAGE" title="SDS-PAGE">SDS-PAGE</a>.<sup id="cite_ref-4" class="reference"><a href="#cite_note-4"><span class="cite-bracket">[</span>4<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-5" class="reference"><a href="#cite_note-5"><span class="cite-bracket">[</span>5<span class="cite-bracket">]</span></a></sup>
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<div class="mw-heading mw-heading2"><h2 id="Eigenschaften">Eigenschaften</h2></div>
<p>SYPRO Orange bindet wie auch sein <a href="Analogon_(Chemie)" title="Analogon (Chemie)">Analogon</a> <a href="SYPRO_Red" title="SYPRO Red">SYPRO Red</a> selektiv an Proteine über 6,5 <a href="Kilodalton" class="mw-redirect" title="Kilodalton">Kilodalton</a> (auch <a href="Glykoprotein" class="mw-redirect" title="Glykoprotein">Glykoproteine</a>) in <a href="Natriumlaurylsulfat" title="Natriumlaurylsulfat">SDS</a>-<a href="Micelle" class="mw-redirect" title="Micelle">Micellen</a>,<sup id="cite_ref-Demchenko_1-2" class="reference"><a href="#cite_note-Demchenko-1"><span class="cite-bracket">[</span>1<span class="cite-bracket">]</span></a></sup> nicht jedoch an <a href="DNA" class="mw-redirect" title="DNA">DNA</a> oder <a href="Lipopolysaccharid" class="mw-redirect" title="Lipopolysaccharid">Lipopolysaccharide</a>.<sup id="cite_ref-Sigma_6-0" class="reference"><a href="#cite_note-Sigma-6"><span class="cite-bracket">[</span>6<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-7" class="reference"><a href="#cite_note-7"><span class="cite-bracket">[</span>7<span class="cite-bracket">]</span></a></sup> SYPRO Orange ist gemäß <a href="Verordnung_(EG)_Nr._1272/2008_(CLP)" title="Verordnung (EG) Nr. 1272/2008 (CLP)">Verordnung (EG) Nr. 1272/2008 (CLP)</a> nicht als gefährlich eingestuft,<sup id="cite_ref-Sigma_6-1" class="reference"><a href="#cite_note-Sigma-6"><span class="cite-bracket">[</span>6<span class="cite-bracket">]</span></a></sup> da seine Toxizität und Mutagenität aufgrund der geringen Produktionsmenge unbekannt ist. Das <a href="Absorption_(Physik)" title="Absorption (Physik)">Absorptionsmaximum</a> liegt bei λ = 470 nm, die Emission ist bei λ = 569 nm maximal.<sup id="cite_ref-8" class="reference"><a href="#cite_note-8"><span class="cite-bracket">[</span>8<span class="cite-bracket">]</span></a></sup> Die Sensitivität bei SDS-PA-Gelen liegt mit SYPRO Orange bei etwa 4–8 ng Protein,<sup id="cite_ref-Sigma_6-2" class="reference"><a href="#cite_note-Sigma-6"><span class="cite-bracket">[</span>6<span class="cite-bracket">]</span></a></sup><sup id="cite_ref-Molecular_Probes_9-0" class="reference"><a href="#cite_note-Molecular_Probes-9"><span class="cite-bracket">[</span>9<span class="cite-bracket">]</span></a></sup> also etwas schlechter als bei der <a href="Silberf%C3%A4rbung" title="Silberfärbung">Silberfärbung</a>. Aufgrund der Fluoreszenz besitzt die SYPRO Orange-Färbung bei einer Mengenabschätzung im Vergleich zur Silberfärbung oder <a href="Coomassie" class="mw-redirect" title="Coomassie">Coomassie</a> einen größeren linearen dynamischen Bereich, d. h. eine bessere Korrelation zwischen der Proteinmenge und der Signalintensität.<sup id="cite_ref-Sigma_6-3" class="reference"><a href="#cite_note-Sigma-6"><span class="cite-bracket">[</span>6<span class="cite-bracket">]</span></a></sup> SYPRO Orange eignet sich nicht zur Färbung von Proteinen auf <a href="Western_Blot" title="Western Blot">Western Blots</a>, <a href="2D-Gelelektrophorese" title="2D-Gelelektrophorese">2D-Gelen</a> oder <a href="Isoelektrische_Fokussierung" title="Isoelektrische Fokussierung">IEF-Gelen</a>.<sup id="cite_ref-Sigma_6-4" class="reference"><a href="#cite_note-Sigma-6"><span class="cite-bracket">[</span>6<span class="cite-bracket">]</span></a></sup> Im Gegensatz zur Silber- und Coomassie-Färbung werden zur Fixierung vor einer Färbung zwar auch Essigsäure, aber keine <a href="Alkohole" title="Alkohole">Alkohole</a> wie <a href="Methanol" title="Methanol">Methanol</a> oder <a href="Ethanol" title="Ethanol">Ethanol</a> verwendet, da SYPRO Orange zur Färbung auf die Anwesenheit von SDS angewiesen ist.<sup id="cite_ref-Molecular_Probes_9-1" class="reference"><a href="#cite_note-Molecular_Probes-9"><span class="cite-bracket">[</span>9<span class="cite-bracket">]</span></a></sup> SYPRO Orange wird als 10 mM Lösung in <a href="Dimethylsulfoxid" title="Dimethylsulfoxid">Dimethylsulfoxid</a> geliefert.<sup id="cite_ref-10" class="reference"><a href="#cite_note-10"><span class="cite-bracket">[</span>10<span class="cite-bracket">]</span></a></sup>
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<div class="mw-heading mw-heading2"><h2 id="Einzelnachweise">Einzelnachweise</h2></div>
<ol class="references">
<li id="cite_note-Demchenko-1"><span class="mw-cite-backlink">↑ <sup><a href="#cite_ref-Demchenko_1-0">a</a></sup> <sup><a href="#cite_ref-Demchenko_1-1">b</a></sup> <sup><a href="#cite_ref-Demchenko_1-2">c</a></sup></span> <span class="reference-text">Aleksandr Petrovich Demchenko: <i>Advanced Fluorescence Reporters in Chemistry and Biology III: Applications in Sensing and Imaging Band 3 von Advanced Fluorescence Reporters in Chemistry and Biology</i>. Springer 2011. ISBN 9783642180354. S. 179ff.</span>
</li>
<li id="cite_note-EP0774122-2"><span class="mw-cite-backlink">↑ <sup><a href="#cite_ref-EP0774122_2-0">a</a></sup> <sup><a href="#cite_ref-EP0774122_2-1">b</a></sup></span> <span class="reference-text"><span class="cite">Patent <a rel="nofollow" class="external text" href="https://worldwide.espacenet.com/publicationDetails/biblio?locale=de_EP&CC=EP&NR=0774122B1&FT=D&KC=B1">EP0774122B1</a>: <i>Protein-Färbung mittels Merocyanin Farbstoffe.</i> Angemeldet am <span style="white-space:nowrap;">20. Mai 1996</span>, veröffentlicht am <span style="white-space:nowrap;">19. August 2009</span>, Anmelder: Molecular Probes Inc, Erfinder: Richard P. Haugland et al.</span><span class="Z3988" title="ctx_ver=Z39.88-2004&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Apatent&rft_id=EP0774122B1&rft.applcc=EP&rft.title=Protein-F%C3%A4rbung+mittels+Merocyanin+Farbstoffe&rft.inventor=Richard+P.+Haugland+et+al&rft.assignee=Molecular+Probes+Inc&rft.appldate=1996-05-20&rft.pubdate=2009-08-19"></span></span>
</li>
<li id="cite_note-3"><span class="mw-cite-backlink"><a href="#cite_ref-3">↑</a></span> <span class="reference-text">T. Kroeger, B. Frieg, T. Zhang, F. K. Hansen, A. Marmann, P. Proksch, L. Nagel-Steger, G. Groth, S. H. Smits, H. Gohlke: <i>EDTA aggregates induce SYPRO orange-based fluorescence in thermal shift assay.</i> In: <i><a href="PLOS_ONE" title="PLOS ONE">PLOS ONE</a>.</i> Band 12, Nummer 5, 2017, S. e0177024, <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.1371/journal.pone.0177024">10.1371/journal.pone.0177024</a></span>, <a class="external mw-magiclink-pmid" rel="nofollow" href="https://www.ncbi.nlm.nih.gov/pubmed/28472107?dopt=Abstract">PMID 28472107</a>, <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5417642/">PMC 5417642</a> (freier Volltext).</span>
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<li id="cite_note-4"><span class="mw-cite-backlink"><a href="#cite_ref-4">↑</a></span> <span class="reference-text">T. H. Steinberg, L. J. Jones, R. P. Haugland, V. L. Singer: <i>SYPRO orange and SYPRO red protein gel stains: one-step fluorescent staining of denaturing gels for detection of nanogram levels of protein.</i> In: <i>Analytical biochemistry.</i> Band 239, Nummer 2, August 1996, S. 223–237, <span class="-print"><a href="Internationale_Standardnummer_f%C3%BCr_fortlaufende_Sammelwerke" title="Internationale Standardnummer für fortlaufende Sammelwerke">ISSN</a> <span style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://zdb-katalog.de/list.xhtml?t=iss%3D%220003-2697%22&key=cql">0003-2697</a></span></span>. <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.1006/abio.1996.0319">10.1006/abio.1996.0319</a></span>. <a class="external mw-magiclink-pmid" rel="nofollow" href="https://www.ncbi.nlm.nih.gov/pubmed/8811914?dopt=Abstract">PMID 8811914</a>.</span>
</li>
<li id="cite_note-5"><span class="mw-cite-backlink"><a href="#cite_ref-5">↑</a></span> <span class="reference-text">S. Gallagher, D. Chakavarti: <i>Staining proteins in gels.</i> In: <i>Journal of visualized experiments : JoVE.</i> Nummer 17, 2008, <span class="-print"><a href="Internationale_Standardnummer_f%C3%BCr_fortlaufende_Sammelwerke" title="Internationale Standardnummer für fortlaufende Sammelwerke">ISSN</a> <span style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://zdb-katalog.de/list.xhtml?t=iss%3D%221940-087X%22&key=cql">1940-087X</a></span></span>. <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.3791/760">10.3791/760</a></span>. <a class="external mw-magiclink-pmid" rel="nofollow" href="https://www.ncbi.nlm.nih.gov/pubmed/19066521?dopt=Abstract">PMID 19066521</a>. <a rel="nofollow" class="external text" href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3253607/">PMC 3253607</a> (freier Volltext).</span>
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<li id="cite_note-Sigma-6"><span class="mw-cite-backlink">↑ <sup><a href="#cite_ref-Sigma_6-0">a</a></sup> <sup><a href="#cite_ref-Sigma_6-1">b</a></sup> <sup><a href="#cite_ref-Sigma_6-2">c</a></sup> <sup><a href="#cite_ref-Sigma_6-3">d</a></sup> <sup><a href="#cite_ref-Sigma_6-4">e</a></sup></span> <span class="reference-text">Datenblatt <span style="font-style:italic;"><a rel="nofollow" class="external text" href="https://www.sigmaaldrich.com/catalog/product/SIGMA/S5692">SYPRO Orange Protein Gel Stain</a></span> bei <a href="Sigma-Aldrich" title="Sigma-Aldrich">Sigma-Aldrich</a>, abgerufen am 28. November 2013 (<a rel="nofollow" class="external text" href="https://www.sigmaaldrich.com/DE/de/sds/SIGMA/S5692?userType=anonymous">PDF</a>).<span class="editoronly" style="display:none;"></span></span>
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<li id="cite_note-7"><span class="mw-cite-backlink"><a href="#cite_ref-7">↑</a></span> <span class="reference-text">T. H. Steinberg, R. P. Haugland, V. L. Singer: <i>Applications of SYPRO orange and SYPRO red protein gel stains.</i> In: <i>Analytical biochemistry.</i> Band 239, Nummer 2, August 1996, S. 238–245, <span class="-print"><a href="Internationale_Standardnummer_f%C3%BCr_fortlaufende_Sammelwerke" title="Internationale Standardnummer für fortlaufende Sammelwerke">ISSN</a> <span style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://zdb-katalog.de/list.xhtml?t=iss%3D%220003-2697%22&key=cql">0003-2697</a></span></span>. <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.1006/abio.1996.0320">10.1006/abio.1996.0320</a></span>. <a class="external mw-magiclink-pmid" rel="nofollow" href="https://www.ncbi.nlm.nih.gov/pubmed/8811917?dopt=Abstract">PMID 8811917</a>.</span>
</li>
<li id="cite_note-8"><span class="mw-cite-backlink"><a href="#cite_ref-8">↑</a></span> <span class="reference-text">Harvey, P. H. & Pagel, M. D. <i>The comparative method in evolutionary biology.</i> Oxford University Press, Oxford (1991).</span>
</li>
<li id="cite_note-Molecular_Probes-9"><span class="mw-cite-backlink">↑ <sup><a href="#cite_ref-Molecular_Probes_9-0">a</a></sup> <sup><a href="#cite_ref-Molecular_Probes_9-1">b</a></sup></span> <span class="reference-text">Molecular Probes / Life Technologies: <i>SYPRO Orange and SYPRO Red Protein Gel Stains.</i> 2003. <a rel="nofollow" class="external text" href="http://tools.lifetechnologies.com/content/sfs/manuals/mp06650.pdf">PDF</a>.</span>
</li>
<li id="cite_note-10"><span class="mw-cite-backlink"><a href="#cite_ref-10">↑</a></span> <span class="reference-text">T. H. Steinberg: <i>Protein gel staining methods: an introduction and overview.</i> In: <i>Methods in enzymology.</i> Band 463, 2009, S. 541–563, <a href="Digital_Object_Identifier" title="Digital Object Identifier">doi</a>:<span class="uri-handle" style="white-space:nowrap"><a rel="nofollow" class="external text" href="https://doi.org/10.1016/S0076-6879%2809%2963031-7">10.1016/S0076-6879(09)63031-7</a></span>, <a class="external mw-magiclink-pmid" rel="nofollow" href="https://www.ncbi.nlm.nih.gov/pubmed/19892191?dopt=Abstract">PMID 19892191</a>.</span>
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