Name: quantitative proteomics using reductive dimethylation for stable isotope labeling
Uploaded: 2014-07-01T14:00:00
Description: Watch this Scientific Journal Video about Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling at JoVE.com

We thank SP Gygi and GM Church for help and guidance. This work was supported by a CNRS chaire d'excellence to ACT.

OPMERKING: Deze methode werd eerder beschreven 12. 1. Eiwitisolatie Bereid 1 mg cellulair eiwit door het lyseren van cellen, bij voorkeur door fysische werkwijzen zoals Franse pers, kralen slaan of sonicatie. Vermijd-lysozym-gemedieerde cellysis omdat het enzym massaspectrometrie metingen zullen beschamen. 2. TCA precipitatie van de eiwitten Voeg 1 volume trichloorazijnzuur (TCA) aan 4 delen eiwit e...

<ol>
	<li>Washburn, M. P., Ulaszek, R., Deciu, C., Schieltz, D. M., Yates III, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+quantitative+proteomic+data+generated+via+multidimensional+protein+identification+technology.">Analysis of quantitative proteomic data generated via multidimensional protein identification technology.</a> Analytical chemistry. 74 (7), 1650-1657 (2002).</li><li>Ong, S. -. E., Blagoev, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable+isotope+labeling+by+amino+acids+in+cell+culture,+SILAC,+as+a+simple+and+accurate+approach+to+expression+proteomics.">Stable isotope labeling by amino acids in cell culture, SILAC, as a simple and accurate approach to expression proteomics.</a> Molecular &amp; cellular proteomics: MCP. 1 (5), 376-386 (2002).</li><li>Gygi, S. P., Rist, B., Gerber, S. A., Turecek, F., Gelb, M. H., Aebersold, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+analysis+of+complex+protein+mixtures+using+isotope-coded+affinity+tags.">Quantitative analysis of complex protein mixtures using isotope-coded affinity tags.</a> Nature biotechnology. 17 (10), 994-999 (1999).</li><li>Ross, P. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiplexed+protein+quantitation+in+Saccharomyces+cerevisiae+using+amine-reactive+isobaric+tagging+reagents.">Multiplexed protein quantitation in Saccharomyces cerevisiae using amine-reactive isobaric tagging reagents.</a> Molecular &amp; cellular proteomics: MCP. 3 (12), 1154-1169 (2004).</li><li>Hsu, J. -. L., Huang, S. -. Y., Chow, N. -. H., Chen, S. -. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stable-isotope+dimethyl+labeling+for+quantitative+proteomics.">Stable-isotope dimethyl labeling for quantitative proteomics.</a> Analytical Chemistry. 75 (24), 6843-6852 (2003).</li><li>Chick, J. M., Haynes, P. A., Molloy, M. P., Bjellqvist, B., Baker, M. S., Len, A. C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+rat+liver+membrane+proteome+using+peptide+immobilized+pH+gradient+isoelectric+focusing.">Characterization of the rat liver membrane proteome using peptide immobilized pH gradient isoelectric focusing.</a> Journal of Proteome Research. 7 (3), 1036-1045 (2008).</li><li>Rappsilber, J., Ishihama, Y., Mann, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stop+and+go+extraction+tips+for+matrix-assisted+laser+desorption/ionization,+nanoelectrospray,+and+LC/MS+sample+pretreatment+in+proteomics.">Stop and go extraction tips for matrix-assisted laser desorption/ionization, nanoelectrospray, and LC/MS sample pretreatment in proteomics.</a> Analytical chemistry. 75 (3), 663-670 (2003).</li><li>Eng, J. K., McCormack, A. L., Yates III, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+approach+to+correlate+tandem+mass+spectral+data+of+peptides+with+amino+acid+sequences+in+a+protein+database.">An approach to correlate tandem mass spectral data of peptides with amino acid sequences in a protein database.</a> Journal of the American Society for Mass Spectrometry. 5 (11), 976-989 (1994).</li><li>Elias, J. E., Gygi, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Target-decoy+search+strategy+for+increased+confidence+in+large-scale+protein+identifications+by+mass+spectrometry.">Target-decoy search strategy for increased confidence in large-scale protein identifications by mass spectrometry.</a> Nature methods. 4 (3), 207-214 (2007).</li><li>Bakalarski, C. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+peptide+abundance+and+dynamic+range+on+stable-isotope-based+quantitative+proteomic+analyses.">The impact of peptide abundance and dynamic range on stable-isotope-based quantitative proteomic analyses.</a> Journal of Proteome Research. 7 (11), 4756-4765 (2008).</li><li>Wilson-Grady, J. T., Haas, W., Gygi, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+comparison+of+the+fasted+and+re-fed+mouse+liver+phosphoproteomes+using+lower+pH+reductive+dimethylation.">Quantitative comparison of the fasted and re-fed mouse liver phosphoproteomes using lower pH reductive dimethylation.</a> Methods (San Diego, Calif.). , (2013).</li><li>Tolonen, A. C., Haas, W., Chilaka, A. C., Aach, J., Gygi, S. P., Church, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Proteome-wide+systems+analysis+of+a+cellulosic+biofuel-producing+microbe.">Proteome-wide systems analysis of a cellulosic biofuel-producing microbe.</a> Molecular Systems Biology. 7, 461 (2011).</li><li>Tolonen, A. C., Chilaka, A. C., Church, G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+gene+inactivation+in+Clostridium+phytofermentans+shows+that+cellulose+degradation+requires+the+family+9+hydrolase+Cphy3367.">Targeted gene inactivation in Clostridium phytofermentans shows that cellulose degradation requires the family 9 hydrolase Cphy3367.</a> Molecular Microbiology. 74 (6), 1300-1313 (2009).</li><li>Munoz, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+quantitative+proteomes+of+human-induced+pluripotent+stem+cells+and+embryonic+stem+cells.">The quantitative proteomes of human-induced pluripotent stem cells and embryonic stem cells.</a> Molecular systems biology. 7, 550 (2011).</li><li>Kovanich, D., Cappadona, S., Raijmakers, R., Mohammed, S., Scholten, A., Heck, A. J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+stable+isotope+dimethyl+labeling+in+quantitative+proteomics.">Applications of stable isotope dimethyl labeling in quantitative proteomics.</a> Analytical and bioanalytical chemistry. 404 (4), 991-1009 (2012).</li><li>McAlister, G. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increasing+the+multiplexing+capacity+of+TMTs+using+reporter+ion+isotopologues+with+isobaric+masses.">Increasing the multiplexing capacity of TMTs using reporter ion isotopologues with isobaric masses.</a> Analytical chemistry. 84 (17), 7469-7478 (2012).</li><li>Boersema, P. J., Aye, T. T., Van Veen, T. A. B., Heck, A. J. R., Mohammed, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Triplex+protein+quantification+based+on+stable+isotope+labeling+by+peptide+dimethylation+applied+to+cell+and+tissue+lysates.">Triplex protein quantification based on stable isotope labeling by peptide dimethylation applied to cell and tissue lysates.</a> Proteomics. 8 (22), 4624-4632 (2008).</li><li>Wu, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Five-plex+isotope+dimethyl+labeling+for+quantitative+proteomics.">Five-plex isotope dimethyl labeling for quantitative proteomics.</a> Chemical communications (Cambridge, England). , (2014).</li><li>Boersema, P. J., Raijmakers, R., Lemeer, S., Mohammed, S., Heck, A. J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiplex+peptide+stable+isotope+dimethyl+labeling+for+quantitative+proteomics.">Multiplex peptide stable isotope dimethyl labeling for quantitative proteomics.</a> Nature Protocols. 4 (4), 484-494 (2009).</li><li>She, Y. -. M., Rosu-Myles, M., Walrond, L., Cyr, T. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+protein+isoforms+in+mesenchymal+stem+cells+by+reductive+dimethylation+of+lysines+in+intact+proteins.">Quantification of protein isoforms in mesenchymal stem cells by reductive dimethylation of lysines in intact proteins.</a> Proteomics. 12 (3), 369-379 (2012).</li><li>Chen, S. -. H., Chen, C. -. R., Chen, S. -. H., Li, D. -. T., Hsu, J. -. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improved+N(&#945;)-acetylated+Peptide+Enrichment+Following+Dimethyl+Labeling+and+SCX.">Improved N(&#945;)-acetylated Peptide Enrichment Following Dimethyl Labeling and SCX.</a> Journal of proteome research. 12 (7), 3277-3287 (2013).</li><li>Sun, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capture+and+Dimethyl+Labeling+of+Glycopeptides+on+Hydrazide+Beads+for+Quantitative+Glycoproteomics+Analysis.">Capture and Dimethyl Labeling of Glycopeptides on Hydrazide Beads for Quantitative Glycoproteomics Analysis.</a> Analytical Chemistry. 84 (20), 8452-8456 (2012).</li></ol>

Een aantal punten maken stabiele isotoop labeling van peptiden met behulp van reductieve dimethylering (Redi etikettering) een aantrekkelijke methode voor kwantitatieve proteomics: goedkoop etikettering reagens (reagentia kost minder dan $ 1 per monster), snelle reactiesnelheid (~ 10 min), de afwezigheid van nevenproducten, hoge reproduceerbaarheid (figuren 3, 4), stabiele reactieproducten, mogelijkheid om een protease te gebruiken en hoge ionisatie-efficiëntie van gelabelde peptiden. Chemische labelin...

Het meten van de concentratie verschillen van veel eiwitten tussen complexe monsters is een centrale uitdaging in proteomics. In toenemende mate wordt dit gedaan door het merken van eiwitten in elk monster met verschillende isotopische labels combineren van de monsters en met massaspectrometrie om concentratieverschillen kwantificeren. Verschillende methoden bestaan ​​voor stabiele isotopische labeling van eiwitten en peptiden. 15 N etikettering 1 en 2 SILAC voeren isotopische labels metabolisch in vivo, terwijl iCAT 3, iTRAQ 4 en vermindering dimethylering 5 commentaar stabiele isotoop labels na eiwitextractie en vertering. Onder deze methoden, is reductieve dimethylering (Redi etikettering) aan populariteit wint als een goedkope en reproduceerbare methode om eiwitconcentratie verschillen in bijna elk type monster te kwantificeren. Redi etikettering omvat peptiden reageren met formaldehyde om een ​​Schiff base, dat vervolgens wordt gereduceerd vormendoor cyanoboorhydride. Deze reactie dimethylates vrije aminogroepen op N-uiteinden en lysinezijketens en monomethylates N-terminale prolines. De hier beschreven protocol methyleert peptiden in monster 1 met een &quot;lichte&quot; label met reagentia met waterstofatomen in hun natuurlijke isotopische distributie en monster 2 met een &quot;zware&quot; label met gedeutereerd formaldehyde en dride (figuur 1). Elke digemethyleerd aminogroep op een peptide resulteert in een massaverschil van 6,0377 Da tussen lichte en zware vormen, die wordt gebruikt om onderscheid te maken tussen de twee vormen met een massaspectrometer. In het bijzonder, zijn ten opzichte van peptide abundanties gekwantificeerd als de verhouding van MS1 ​​gewonnen ionenchromatogram gebieden (MS1 piekoppervlakte ratio) van lichte en zware uitvoering voor elk peptide ionenpaar. De relatieve abundantie van een eiwit wordt berekend als de mediaan MS1 ​​piekoppervlak verhouding tussen alle peptiden in het proteïne. In dit rapport beschrijven we een robuust protocol voor gedraging Redi markeringsexperimenten door LC-MS/MS dat omgekeerde fase peptide vaste fase extractie op-kolom Redi etikettering peptide fractionering door basische pH omgekeerde fase (BPRP) chromatografie en zuivering van peptide mengsels met StageTips (figuur 2) omvat . We bespreken de voordelen en beperkingen van het gebruik van Redi etikettering voor kwantitatieve proteomics.

<table border="0" cellpadding="0" cellspacing="0" width="973">
	<tbody>
		<tr height="19">
			<td height="19" style="height:19px;width:377px;">trichloroacetic acid</td>
			<td style="width:141px;">Sigma-Aldrich</td>
			<td style="width:136px;">T9159</td>
			<td style="width:319px;">protein precipitation</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">acetone</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">650501</td>
			<td>protein precipitation</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">Sodium dodecyl sulfate</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">71736</td>
			<td>denature, reduce protein</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">sodium hydroxide</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">S8045</td>
			<td>denature, reduce protein</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">DL-Dithiothreitol</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">43816</td>
			<td>denature, reduce, alkylate protein</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">protease Inhibitor Complete Mini Cocktail</td>
			<td>Roche</td>
			<td>4693124001</td>
			<td>denature, reduce protein</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">iodoacetamide</td>
			<td>Sigma-Aldrich</td>
			<td>I6125</td>
			<td>alkylate protein</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">HEPES</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">H7523</td>
			<td>resuspend, extract, label protein</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">calcium chloride</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">C5670</td>
			<td>resuspend protein</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:377px;">Lysyl Endoprotease</td>
			<td>Wako Chemicals</td>
			<td style="width:136px;">129-02541</td>
			<td>protein digestion</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">sequencing grade trypsin</td>
			<td>Promega</td>
			<td style="width:136px;">V5111</td>
			<td>protein digestion</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">acetic acid</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">320099</td>
			<td>protein digestion</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">trifluoroacetic acid</td>
			<td>Sigma-Aldrich</td>
			<td>299537</td>
			<td>Reversed-phase peptide extraction</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">tC18 Sep-Pak C18 cartridges</td>
			<td>Waters</td>
			<td>WAT054960</td>
			<td>Reversed-phase peptide extraction</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">extraction manifold</td>
			<td>Waters</td>
			<td>WAT200609</td>
			<td>Reversed-phase peptide extraction</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">acetonitrile</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">14261</td>
			<td>various</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">formaldehyde</td>
			<td>Sigma-Aldrich</td>
			<td>252549</td>
			<td>&#8220;light&#8221; peptide labeling</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">cyanoborohydride</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">71435</td>
			<td>&#8220;light&#8221; peptide labeling</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">deuterated formaldehyde</td>
			<td>Sigma-Aldrich</td>
			<td>492620</td>
			<td>&#8220;heavy&#8221; peptide labeling</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">sodium cyanoborodeuteride</td>
			<td>CDN isotopes</td>
			<td style="width:136px;">D-1797</td>
			<td>&#8220;heavy&#8221; peptide labeling</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">MES</td>
			<td>Sigma-Aldrich</td>
			<td>M3671</td>
			<td>peptide labeling</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">C18-HPLC column (4.6 x 250 mm, 5 &#181;m particle size)</td>
			<td>Agilent</td>
			<td style="width:136px;">770450-902</td>
			<td>basic pH reversed-phase chromatography</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">formic acid</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">399388</td>
			<td>various</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">C18 Empore Disks</td>
			<td>3M</td>
			<td style="width:136px;">14-386-3&#160;</td>
			<td>STAGE tips</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;">methanol</td>
			<td>Sigma-Aldrich</td>
			<td style="width:136px;">494437</td>
			<td>STAGE tips</td>
		</tr>
	</tbody>
</table>

quantitative proteomics using reductive dimethylation for stable isotope labeling

Stabiele isotopen van peptiden door reductieve dimethylering (Redi labeling) is een methode om nauwkeurig kwantificeren eiwitexpressie verschillen tussen monsters met behulp van massaspectrometrie. Redi etikettering wordt uitgevoerd met behulp van gewone (licht) of gedeutereerde (zware) vormen van formaldehyde en natriumcyaanboriumhydride twee methylgroepen toevoegen aan elke vrije amine. Hier laten we zien een robuust protocol voor Redi etikettering en kwantitatieve vergelijking van complexe eiwitmengsels. Eiwitmonsters ter vergelijking worden gedigesteerd in peptiden gelabeld ofwel lichte of zware methyl markeringen, gemengd en samen geanalyseerd met LC-MS/MS dragen. Relatieve dichtheden eiwit gekwantificeerd door vergelijking van de ionenchromatogram piekoppervlakten van zware en lichte gemerkte versies van de samenstellende peptide geëxtraheerd uit de volledige MS-spectra. De hier beschreven methode is inclusief monstervoorbereiding met reversed phase vaste-fase-extractie, on-column Redi etikettering van peptiden, peptide fractionering door basische pH reversed-fase (BPRP) chromatografie, en StageTip peptide zuivering. We bespreken de voordelen en beperkingen van Redi etikettering met betrekking tot andere methoden voor een stabiele isotoop oprichting. We belichten nieuwe toepassingen met Redi labeling als een snelle, goedkope en nauwkeurige methode om eiwit abondantie vergelijken in bijna elk soort monster.

We evalueerden de nauwkeurigheid, precisie en reproduceerbaarheid van Redi labeling met Saccharomyces cerevisiae en Clostridium phytofermentans gehele cellysaten. We hebben eerst gekwantificeerd de Redi etikettering efficiëntie van een mix van C. phytofermentans eiwitlysaten uit cellulose (zware gelabeld, H) en glucose (licht label, L) culturen. Wanneer gefilterd om een ​​1% peptide valse ontdekking tarief, dit monster bevatte 11.194 unieke peptide sequenties met een 98% Redi etikettering...

Watch this Scientific Journal Video about Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling at JoVE.com

Quantitative Proteomics Using Reductive Dimethylation for Stable Isotope Labeling

Kwantitatieve Proteomics behulp Reductieve dimethylering voor stabiele isotopen Labeling

Stabiele isotoop labeling van peptiden door reductieve dimethylering (Redi etikettering) is een snelle, goedkope strategie voor nauwkeurige massa-spectrometrie gebaseerde kwantitatieve proteomics. Hier laten we een robuuste werkwijze voor de bereiding en analyse van eiwitmengsels met de Redi aanpak die kan worden toegepast op vrijwel elk type monster.

Stable isotope labeling of peptides by reductive dimethylation (ReDi labeling) is a method to accurately quantify protein expression differences between ...

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CN-GELFrEE - Clear Native Gel-eluted Liquid Fraction Entrapment Electrophoresis

CN-GELFrEE - Clear Inheems Gel-uitgewassen vloeibare fractie Entrapment elektroforese

Protein complexes perform an array of crucial cellular functions. Elucidating their non-covalent interactions and dynamics is paramount for understanding ...

A Quantitative Glycomics and Proteomics Combined Purification Strategy

Een kwantitatieve Glycomics en Proteomics Combined Purification Strategy

There is a growing desire in the biological and clinical sciences to integrate and correlate multiple classes of biomolecules to unravel biology, define ...

Standards for Quantitative Metalloproteomic Analysis Using Size Exclusion ICP-MS

Normen voor Quantitative Metalloproteomic analyse met behulp van Size Exclusion ICP-MS

Metals are essential for protein function as cofactors to catalyze chemical reactions. Disruption of metal homeostasis is implicated in a number of ...

Highly Stable, Functional Hairy Nanoparticles and Biopolymers from Wood Fibers: Towards Sustainable Nanotechnology

Sterk Stabiel, Functioneel Hairy Nanodeeltjes en Biopolymeren uit houtvezels: Towards Sustainable Nanotechnologie

Nanoparticles, as one of the key materials in nanotechnology and nanomedicine, have gained significant importance during the past decade. While ...

Peptide and Protein Quantification Using Automated Immuno-MALDI (iMALDI)

Peptide and Protein Quantification Using Automated Immuno-MALDI iMALDI

Peptide en proteïne kwantificering met behulp van geautomatiseerde Immuno-MALDI (iMALDI)

Mass spectrometry (MS) is one of the most commonly used technologies for quantifying proteins in complex samples, with excellent assay specificity as a ...

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

Snelle nanosonde signaal versterking door In Situ gouden Nanoparticle synthese

The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity ...