Name: peptide and protein quantification using automated immuno-maldi (imaldi)
Uploaded: 2017-08-18T12:30:00
Description: Watch this Scientific Journal Video about Peptide and Protein Quantification Using Automated Immuno-MALDI iMALDI at JoVE.com

我们感谢金融支持从加拿大基因和基因组不列颠哥伦比亚省 (204PRO) 的操作和技术发展 (214PRO) 通过基因组创新网络 (GIN)。我们感谢研究所的麦吉尔大学健康中心拍摄 MALDI TOF 仪用于药物发现平台。H.L.是感谢支持从国家科学和工程研究理事会的加拿大 (NSERC) 博士后。C.H.B 非常感谢支持从前缘养老基金 （叶）。C.H.B.是感谢支持从在麦吉尔大学 （加拿大，魁北克，蒙特利尔） 分子肿瘤学的西格尔麦吉尔椅子。M.X.C.和 C.H.B.都很感激支持沃伦 Y.索珀慈善信托基金和阿尔文 · 西格尔家族基金会往犹太医院 （蒙特利尔，魁北克，加拿大）。

试剂如下所述的数额基于 20 病人血浆样品的测量。议定书 》 提出了以下如下准则的维多利亚大学 &#39; s 人类研究伦理委员会。
 1.昂代我人血浆中 <ol><li>解冻血浆样品 (&#8805; 200 微升) 在一个房间里温度水浴 5 分钟，然后把样品直到完全融化的冰上。</li>
	<li>每个样品的浓度手动来分离 1.1 毫升深孔板 （样板） 井和离心 10 分钟在 2 ° C 和 1278 x g.离心机板转移 200 微升</li>
	<li...

<ol>
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Proteomics. 107, 24-30 (2014).</li><li>Landegren, U., 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=Opportunities+for+Sensitive+Plasma+Proteome+Analysis.">Opportunities for Sensitive Plasma Proteome Analysis.</a> Anal. Chem. 84 (4), 1824-1830 (2012).</li><li>Guo, A., 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=Immunoaffinity+Enrichment+and+Mass+Spectrometry+Analysis+of+Protein+Methylation.">Immunoaffinity Enrichment and Mass Spectrometry Analysis of Protein Methylation.</a> Mol. &amp; Cell. Proteomics. 13 (1), 372-387 (2014).</li><li>Florentinus-Mefailoski, A., Soosaipillai, A., Dufresne, J., Diamandis, E. P., Marshall, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+enzyme-linked+immuno-mass+spectrometric+assay+with+the+substrate+adenosine+monophosphate.">An enzyme-linked immuno-mass spectrometric assay with the substrate adenosine monophosphate.</a> Anal. Bioanal. Chem. 407 (4), 1119-1130 (2015).</li><li>Fredolini, 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=Immunocapture+strategies+in+translational+proteomics.">Immunocapture strategies in translational proteomics.</a> Expert Rev Proteomics. 13 (1), 83-98 (2016).</li><li>Tran, J. 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=Automated+Affinity+Capture+and+On-Tip+Digestion+to+Accurately+Quantitate+in+Vivo+Deamidation+of+Therapeutic+Antibodies.">Automated Affinity Capture and On-Tip Digestion to Accurately Quantitate in Vivo Deamidation of Therapeutic Antibodies.</a> Anal. Chem. , (2016).</li><li>Razavi, M., Leigh Anderson, N., Pope, M. E., Yip, R., Pearson, T. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High+precision+quantification+of+human+plasma+proteins+using+the+automated+SISCAPA+Immuno-MS+workflow.">High precision quantification of human plasma proteins using the automated SISCAPA Immuno-MS workflow.</a> N. Biotechnol. 33 (5 Part A), 494-502 (2016).</li><li>Razavi, M., 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=High-Throughput+SISCAPA+Quantitation+of+Peptides+from+Human+Plasma+Digests+by+Ultrafast,+Liquid+Chromatography-Free+Mass+Spectrometry.">High-Throughput SISCAPA Quantitation of Peptides from Human Plasma Digests by Ultrafast, Liquid Chromatography-Free Mass Spectrometry.</a> J. Proteome Res. 11 (12), 5642-5649 (2012).</li><li>Anderson, N. 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=SISCAPA+Peptide+Enrichment+on+Magnetic+Beads+Using+an+In-line+Bead+Trap+Device.">SISCAPA Peptide Enrichment on Magnetic Beads Using an In-line Bead Trap Device.</a> Mol. &amp; Cell. Proteomics. 8 (5), 995-1005 (2009).</li><li>Parker, C. E., Pearson, T. W., Anderson, N. L., Borchers, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mass-spectrometry-based+clinical+proteomics+-+a+review+and+prospective.">Mass-spectrometry-based clinical proteomics - a review and prospective.</a> The Analyst. 135 (8), 1830-1838 (2010).</li><li>Reid, J. D., Holmes, D. T., Mason, D. R., Shah, B., Borchers, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Towards+the+Development+of+an+Immuno+MALDI+(iMALDI)+Mass+Spectrometry+Assay+for+the+Diagnosis+of+Hypertension.">Towards the Development of an Immuno MALDI (iMALDI) Mass Spectrometry Assay for the Diagnosis of Hypertension.</a> J. Am. Soc. Mass Spectrom. 21 (10), 1680-1686 (2010).</li><li>Mason, D. R., Reid, J. D., Camenzind, A. G., Holmes, D. T., Borchers, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Duplexed+iMALDI+for+the+detection+of+angiotensin+I+and+angiotensin+II.">Duplexed iMALDI for the detection of angiotensin I and angiotensin II.</a> Methods. 56 (2), 213-222 (2012).</li><li>Camenzind, A. G., vander Gugten, J. G., Popp, R., Holmes, D. T., Borchers, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+evaluation+of+an+immuno-MALDI+(iMALDI)+assay+for+angiotensin+I+and+the+diagnosis+of+secondary+hypertension.">Development and evaluation of an immuno-MALDI (iMALDI) assay for angiotensin I and the diagnosis of secondary hypertension.</a> Clin. Proteomics. 10 (1), 20 (2013).</li><li>Jiang, 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=Development+of+an+immuno+tandem+mass+spectrometry+(iMALDI)+assay+for+EGFR+diagnosis.">Development of an immuno tandem mass spectrometry (iMALDI) assay for EGFR diagnosis.</a> Proteomics Clin Appl. 1, (2007).</li><li>Jiang, J., Parker, C. E., Fuller, J. R., Kawula, T. H., Borchers, C. 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M., 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=Automated+screening+of+monoclonal+antibodies+for+SISCAPA+assays+using+a+magnetic+bead+processor+and+liquid+chromatography-selected+reaction+monitoring-mass+spectrometry.">Automated screening of monoclonal antibodies for SISCAPA assays using a magnetic bead processor and liquid chromatography-selected reaction monitoring-mass spectrometry.</a> J Immunol Methods. 353 (1-2), 49-61 (2010).</li><li>Borchers, C. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+of+quantitation+and+identification+of+peptides+and+proteins.">Methods of quantitation and identification of peptides and proteins.</a> US patent. , (2010).</li><li>Borchers, C. 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相比传统的基于 MS 蛋白质定量，iMALDI 使用抗体来丰富分析物和净化他们从复杂的样品，因此使它能够量化的蛋白质或多肽在低浓度时。IMALDI 协议的关键一步是目标肽免疫富集。为此目的，应选择具有高特异性和亲和力抗体。在 SISCAPA，报道在 10-9米或更好的抗体亲和力将需要实现高灵敏度低吴/毫升21。此外，与高的抗体结合能力的珠子是首选最优浓缩。
<p class="jove_co...

质谱技术已经成为不可或缺的工具，在定量蛋白质组学。质谱分析可以确定目标蛋白质或多肽的群众，因此所得的分析物信号可以高度特异性相比于免疫分析。两种电离方法，电喷雾串联质谱和 MALDI，常被用于检测蛋白质和多肽1，，23，4。在基于 MS 蛋白质定量的主要挑战在于复杂样品在吴/毫升或 pg/mL 的浓度在高丰度蛋白低丰度蛋白的检测及血浆中发现很多候选人蛋白质生物标志物在这范围之内5。这个问题很大程度上造成的人类蛋白质组6复杂性与本质上宽动态范围。
为了克服这些检测，免疫色谱-质谱方法已充实目标蛋白质或多肽从样品溶液在固体的表面，紧接着洗脱物和 MS 测量7，8,9,10.通过免疫浓缩物都从复杂样品纯化，因此从其他分子离子抑制影响降至最低。各种固体支架间磁珠是目前使用最广泛那样高的抗体结合能力的优势和易用性的处理。磁珠与不同的 functionalizations 和大小有被开发和商业化免疫沉淀实验。到目前为止，免疫浓缩制成的小珠已经配套与电喷雾电离 (ESI) 和 MALDI 质谱蛋白质和多肽的测量。稳定同位素标准和捕获由抗多肽抗体 (SISCAPA) 技术，在样品中的蛋白质消化，紧接着孵化与抗体包裹小珠免疫富集。在"古典"SISCAPA，捕获的 proteotypic 多肽是从珠、 洗脱，由液相色谱-质谱 (LC-MS)，或直接输注 ESI 多反应监测-质谱 (ESI-MRM-MS)11，12来衡量。免疫富集 MRM 检测灵敏度提高 3-4 个数量级，达到低吴/mL 范围13。
与电喷雾质谱相比，质谱速度更快，并且不涉及清洗和重新平衡的 LC 列所以没有携带量和污染的问题，使它更加适合高通量研究14。免疫 MALDI 技术得到了我们的实验室要结合质谱免疫富集的敏感和具体量化的多肽和蛋白质 （基于 proteotypic 多肽定量）15，16 ，17。免疫富集，珠 MALDI 靶板上沉积、 矩阵解的方法添加到珠子，和干燥后板是准备 TOF 质谱分析。从珠肽洗脱不执行作为一个单独的步骤，但亲和绑定物洗脱 MALDI 基质解决方案时将其添加到珠点，从而简化样品制备和样品损失最小化。IMALDI 技术已被应用于各种应用程序18，19，最近经过自动化和用于测量血管紧张素我 (和我) 测定血浆肾素活性 (PRA)20。本议定书将演示用于执行自动化的 iMALDI 测定的过程。以 PRA 测定为例，间天 CVs 的少于 10%已通过自动化，与每一天20744 样品的检测能力。
IMALDI PRA 测定表明这份手稿在不需要蛋白质的消化，作为靶分子 (Ang 我) 是一种肽 1295.7 大分子量。在哪里是必要的蛋白质的消化和肽用作完整的蛋白质代孕其他试剂，选定的肽对 iMALDI 应该是独特和特定于目标蛋白，与大量超过 800 的大，所以它可以很容易区分 c从 MALDI 基质溶液的化学噪声。抗多肽抗体所需的多肽免疫富集。IMALDI 试验测量 PRA 的协议包括四个步骤： 1) 昂代我在人血浆;2） 免疫富集昂我使用抗体包被珠;3） 珠转交 MALDI 靶板和添加矩阵解法;和 4） 信号采集20名 MALDI TOF MS 和数据分析。

<table width="892">
	<tbody>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Healthy control human plasma</td>
			<td class="xl63" style="width:134pt" width="178">Bioreclamation</td>
			<td class="xl68" style="width:89pt" width="119">HMPLEDTA2</td>
			<td class="xl66" style="width:242pt" width="323"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Ammonium bicarbonate</td>
			<td class="xl63" style="width:134pt" width="178">Sigma Aldrich</td>
			<td class="xl69">09830</td>
			<td class="xl66"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Ammonium citrate dibasic</td>
			<td class="xl63" style="width:134pt" width="178">Sigma Aldrich</td>
			<td class="xl68" style="width:89pt" width="119">09833</td>
			<td class="xl66"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">CHAPS (&#62;=98%)</td>
			<td class="xl63" style="width:134pt" width="178">Sigma Aldrich</td>
			<td class="xl68" style="width:89pt" width="119">C9426</td>
			<td class="xl66"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Albumin from chicken egg white (&#62;98%)</td>
			<td class="xl63" style="width:134pt" width="178">Sigma Aldrich</td>
			<td class="xl68" style="width:89pt" width="119">A5503</td>
			<td class="xl66"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Ethylenediaminetetraacetic acid</td>
			<td class="xl63" style="width:134pt" width="178">Sigma Aldrich</td>
			<td class="xl68" style="width:89pt" width="119">EDS</td>
			<td class="xl66"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Alpha-cyano-4-hydroxycinnamic acid</td>
			<td class="xl63" style="width:134pt" width="178">Sigma Aldrich</td>
			<td class="xl68" style="width:89pt" width="119">70990</td>
			<td class="xl66"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Phosphate buffered saline</td>
			<td class="xl63" style="width:134pt" width="178">Sigma Aldrich</td>
			<td class="xl68" style="width:89pt" width="119">P4417</td>
			<td class="xl66"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Phenylmethanesulfonyl fluoride</td>
			<td class="xl63" style="width:134pt" width="178">Sigma Aldrich</td>
			<td class="xl68" style="width:89pt" width="119">78830</td>
			<td class="xl66"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Trifluoroacetic acid</td>
			<td class="xl63" style="width:134pt" width="178">Thermo Fisher Scientific</td>
			<td class="xl68" style="width:89pt" width="119">85172</td>
			<td class="xl66">LC-MS grade</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">acetonitrile</td>
			<td class="xl63" style="width:134pt" width="178">Fluka</td>
			<td class="xl68" style="width:89pt" width="119">34967</td>
			<td class="xl66">LC-MS grade</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">water</td>
			<td class="xl63" style="width:134pt" width="178">Fluka</td>
			<td class="xl68" style="width:89pt" width="119">39253</td>
			<td class="xl66">LC-MS grade</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">acetic acid</td>
			<td class="xl63" style="width:134pt" width="178">Fluka</td>
			<td class="xl68" style="width:89pt" width="119">320099</td>
			<td class="xl66">LC-MS grade</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Tris(hydroxymethyl)aminomethane</td>
			<td class="xl63" style="width:134pt" width="178">Roche Diagnostics</td>
			<td class="xl71">3118169001</td>
			<td class="xl66"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Dynabeads Protein G magnetic beads</td>
			<td class="xl63" style="width:134pt" width="178">Thermo Fisher Scientific</td>
			<td class="xl68" style="width:89pt" width="119">10003D</td>
			<td class="xl66">2.8 &#956;m, 30 mg/mL</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">anti-Ang I goat polyclonal antibody</td>
			<td class="xl63" style="width:134pt" width="178">Santa Cruz Biotechnology</td>
			<td class="xl68" style="width:89pt" width="119">sc-7419</td>
			<td class="xl66"></td>
		</tr>
		<tr height="84" style="height:63.0pt">
			<td class="xl63" height="84" style="width:204pt;height:63.0pt" width="272">Nat and SIS Ang I</td>
			<td class="xl63" style="width:134pt" width="178">synthesized at the University of Victoria-Genome BC Proteomics Centre</td>
			<td class="xl68" style="width:89pt" width="119"></td>
			<td class="xl66"></td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Automated liquid handling system</td>
			<td class="xl63" style="width:134pt" width="178">Agilent</td>
			<td class="xl68" style="width:89pt" width="119">16050-102</td>
			<td class="xl66">Agilent Bravo robotic workstation</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Magnet</td>
			<td class="xl63" style="width:134pt" width="178">Thermo Fisher Scientific</td>
			<td class="xl68" style="width:89pt" width="119">12321D</td>
			<td class="xl66">Invitrogen DynaMag-2 magnet</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Tube rotator</td>
			<td class="xl63" style="width:134pt" width="178">Theromo Scientific</td>
			<td class="xl68" style="width:89pt" width="119">400110Q</td>
			<td class="xl66">Labquake Tube Rotator</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Magnet</td>
			<td class="xl63" style="width:134pt" width="178">Thermo Fisher Scientific</td>
			<td class="xl68" style="width:89pt" width="119">12027</td>
			<td class="xl66">DynaMag-96 side skirted magnet</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">Magnet</td>
			<td class="xl63" style="width:134pt" width="178">VP Scientific</td>
			<td class="xl68" style="width:89pt" width="119">771RM-1</td>
			<td class="xl66">used to pull the beads to the bottom of the well</td>
		</tr>
		<tr height="21" style="height:15.75pt">
			<td class="xl63" height="21" style="width:204pt;height:15.75pt" width="272">MALDI-TOF</td>
			<td class="xl63" style="width:134pt" width="178">Bruker</td>
			<td class="xl68" style="width:89pt" width="119"></td>
			<td class="xl66">Bruker Microflex LRF instrument</td>
		</tr>
	</tbody>
</table>

peptide and protein quantification using automated immuno-maldi (imaldi)

质谱 (MS) 是量化复杂样品中的蛋白质与优秀测定特异性由于每个目标分子质量电荷比直接检测的最常用的技术之一。然而，基于 MS 的蛋白质组学，像大多数其他分析技术，有偏向于测量高丰度分析物，所以它难实现检测限制的低吴/毫升或 pg/mL 的复杂样品和这许多的浓度范围是疾病相关蛋白等人血浆的关键问题之一。为协助低丰度分析物的检测，免疫富集，只纳入测定浓缩和纯化前 MS 测量，显著提高了检测灵敏度分析物。在这项工作，免疫-Matrix-Assisted 激光解吸电离 (iMALDI) 技术是提出了一种定量的蛋白质和肽的关键问题之一，基于免疫浓缩制成的小珠，紧接着 MALDI 质谱测量不事先洗脱。抗多肽抗体是对磁珠，官能化和培养与样品。洗完后，珠子直接转嫁 MALDI 靶板和信号测定 (MALDI Tof) 仪器 MALDI 时间后矩阵解的方法已被应用于珠。样品制备程序简化相比其他免疫色谱-质谱测定和 MALDI 测量是快速。整个样品制备自动化与液体处理系统，以提高的测定的重现性和更高的吞吐量。在这篇文章，iMALDI 法用于确定肽血管紧张素我 (Ang 我) 用于临床作为读出的血浆肾素活性的筛选原发性醛固酮增多 (PA) 的血浆中的浓度。

李安的测量自动化的 iMALDI 程序我是图 1中所示。关于抗肽磁珠，丰富了目标肽 （内源性肽或从消化蛋白质肽） 和珠子然后转给 MALDI 测量的靶板。整个过程就简化相比需要更多肽洗脱步骤其他免疫色谱-质谱技术。IMALDI 检测自动化允许高通量分析的大量样品间天变异系数 (CV 10 20以下)。代表性的光谱测定人血浆样品中的我<strong class=...

Watch this Scientific Journal Video about Peptide and Protein Quantification Using Automated Immuno-MALDI iMALDI at JoVE.com

Peptide and Protein Quantification Using Automated Immuno-MALDI iMALDI

提出了在使用自动的免疫-MALDI (iMALDI) 技术的复杂生物体液蛋白质定量的协议。

多肽和蛋白质量化使用自动免疫-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 ...

Research

JoVE Journal

Chemistry

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

Production of Disulfide-stabilized Transmembrane Peptide Complexes for Structural Studies

生产二硫键稳定的跨膜肽复合物的结构研究

Physical  interactions among the lipid-embedded alpha-helical domains of membrane  proteins play a crucial role in folding and assembly of membrane ...

科研

化学

Multiplexed Fluorescent Microarray for Human Salivary Protein Analysis Using Polymer Microspheres and Fiber-optic Bundles

复用荧光微阵列人类唾液蛋白质分析用高分子微球和光纤束

Herein, we describe a protocol for simultaneously measuring six proteins in saliva using a fiber-optic microsphere-based antibody array. The immuno-array ...

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR

Copper (I) binding by metallochaperone transport proteins prevents copper oxidation and release of the toxic ions that may participate in harmful redox ...

Analyzing Protein Dynamics Using Hydrogen Exchange Mass Spectrometry

使用氢交换质谱分析蛋白质动力学

All cellular processes depend on the functionality of proteins. Although the functionality of a given protein is the direct consequence of its unique ...

Split-and-pool Synthesis and Characterization of Peptide Tertiary Amide Library

分裂和池合成及肽叔酰胺图书馆表征

Peptidomimetics are great sources of protein ligands. The oligomeric nature of these compounds enables us to access large synthetic libraries on solid ...

Development of a Backbone Cyclic Peptide Library as Potential Antiparasitic Therapeutics Using Microwave Irradiation

一个主干环肽库作为潜在的抗寄生虫治疗微波辐射的发展

Protein-protein interactions (PPIs) are intimately involved in almost all biological processes and are linked to many human diseases. Therefore, there is ...

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

CN-GELFrEE  - 透明天然凝胶洗脱液体馏分偷天陷阱电泳

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

Characterization, Quantification and Compound-specific Isotopic Analysis of Pyrogenic Carbon Using Benzene Polycarboxylic Acids (BPCA)

Characterization, Quantification and Compound-specific Isotopic Analysis of Pyrogenic Carbon Using Benzene Polycarboxylic Acids BPCA

表征，气相炭用苯聚羧酸的定量和特定化合物同位素分析（BPCA）

Fire-derived, pyrogenic carbon (PyC), sometimes called black carbon (BC), is the carbonaceous solid residue of biomass and fossil fuel combustion, such as ...

Capillary Electrophoresis to Monitor Peptide Grafting onto Chitosan Films in Real Time

毛细管电泳监视肽接枝壳聚糖膜实时

Free-solution capillary electrophoresis (CE) separates analytes, generally charged compounds in solution through the application of an electric field. ...

In Situ Detection and Single Cell Quantification of Metal Oxide Nanoparticles Using Nuclear Microprobe Analysis

In Situ Detection and Single Cell Quantification of Metal Oxide Nanoparticles Using Nuclear Microprobe Analysis

原位金属氧化物纳米粒子的核探针检测与单细胞定量研究

Micro-analytical techniques based on chemical element imaging enable the localization and quantification of chemical composition at the cellular level. ...