Name: calibration-free in vitro quantification of protein homo-oligomerization using commercial instrumentation and free, open source brightness analysis software
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Description: Watch this Scientific Journal Video about Calibration-free In Vitro Quantification of Protein Homo-oligomerization Using Commercial Instrumentation and Free, Open Source Brightness Analysis Software a

这项工作得到了威康信托基金 105278/Z/14/2 对 R.N. 的支持。威康人类遗传学信托中心由威康信托核心奖 203852/Z/16/2 资助。校准小组的工作由英国癌症研究 (C20724/A14414) 和欧洲研究理事会根据欧洲联盟2020研究和创新方案赠款647278提供支持。

1. FKBP12 F36V-mVenus 纯化
<ol><li>转换 (DE3) pLysS 细胞与 pET22b 向量包含 monomerized 人类 FKBP12F36V12和 N 终端 His6 和 mVenus 标签 (向量可根据要求提供)。板细胞到 LB 琼脂补充50µg/毫升氨苄西林和34µg/毫升氯霉素。</li>
	<li>转移转化成的菌落为100毫升磅起始培养和增长 16-20 小时在37°c 与震动。</li>
	<li>稀密启动培养 (OD600 > 1) 1:100 在 LB 介质 (2 x 500 毫升批次) 和成长为 2-3 小时到 OD600 = 0.6-0.8 (37...

<ol>
	<li>Voith von Voithenberg, L., Lamb, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+pair+forster+resonance+energy+transfer:+A+versatile+tool+to+investigate+protein+conformational+dynamics.">Single pair forster resonance energy transfer: A versatile tool to investigate protein conformational dynamics.</a> BioEssays. 40, (2018).</li><li>Padilla-Parra, S., Auduge, N., Coppey-Moisan, M., Tramier, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+FRET+analysis+by+fast+acquisition+time+domain+FLIM+at+high+spatial+resolution+in+living+cells.">Quantitative FRET analysis by fast acquisition time domain FLIM at high spatial resolution in living cells.</a> Biophysical Journal. 95, 2976-2988 (2008).</li><li>Padilla-Parra, S., Auduge, N., Coppey-Moisan, M., Tramier, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dual-color+fluorescence+lifetime+correlation+spectroscopy+to+quantify+protein-protein+interactions+in+live+cell.">Dual-color fluorescence lifetime correlation spectroscopy to quantify protein-protein interactions in live cell.</a> Microscopy Research and Technique. 74, 788-793 (2011).</li><li>Muller, J. D., Chen, Y., Gratton, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence+correlation+spectroscopy.">Fluorescence correlation spectroscopy.</a> Methods in Enzymology. 361, 69-92 (2003).</li><li>Tramier, M., Coppey-Moisan, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence+anisotropy+imaging+microscopy+for+homo-FRET+in+living+cells.">Fluorescence anisotropy imaging microscopy for homo-FRET in living cells.</a> Methods in Cell Biology. 85, 395-414 (2008).</li><li>Digman, M. A., Dalal, R., Horwitz, A. F., Gratton, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+the+number+of+molecules+and+brightness+in+the+laser+scanning+microscope.">Mapping the number of molecules and brightness in the laser scanning microscope.</a> Biophysical Journal. 94, 2320-2332 (2008).</li><li>Nolan, R., Iliopoulou, M., Alvarez, L., Padilla-Parra, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Detecting+protein+aggregation+and+interaction+in+live+cells:+A+guide+to+number+and+brightness.">Detecting protein aggregation and interaction in live cells: A guide to number and brightness.</a> Methods. , (2017).</li><li>Dalal, R. B., Digman, M. A., Horwitz, A. F., Vetri, V., Gratton, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Determination+of+particle+number+and+brightness+using+a+laser+scanning+confocal+microscope+operating+in+the+analog+mode.">Determination of particle number and brightness using a laser scanning confocal microscope operating in the analog mode.</a> Microscopy Research and Technique. 71, 69-81 (2008).</li><li>Nolan, R., 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=nandb-number+and+brightness+in+R+with+a+novel+automatic+detrending+algorithm.">nandb-number and brightness in R with a novel automatic detrending algorithm.</a> Bioinformatics. , (2017).</li><li>Hur, K. H., 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=Quantitative+Measurement+of+Brightness+from+Living+Cells+in+the+Presence+of+Photodepletion.">Quantitative Measurement of Brightness from Living Cells in the Presence of Photodepletion.</a> PLoS One. 9, (2014).</li><li>Amara, J. F., 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=A+versatile+synthetic+dimerizer+for+the+regulation+of+protein-protein+interactions.">A versatile synthetic dimerizer for the regulation of protein-protein interactions.</a> Proceedings of the National Academy of Sciences of the United States of America. 94, 10618-10623 (1997).</li><li>Clackson, T., 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=Redesigning+an+FKBP-ligand+interface+to+generate+chemical+dimerizers+with+novel+specificity.">Redesigning an FKBP-ligand interface to generate chemical dimerizers with novel specificity.</a> Proceedings of the National Academy of Sciences of the United States of America. 95, 10437-10442 (1998).</li><li>Rollins, C. T., 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=A+ligand-reversible+dimerization+system+for+controlling+protein-protein+interactions.">A ligand-reversible dimerization system for controlling protein-protein interactions.</a> Proceedings of the National Academy of Sciences of the United States of America. 97, 7096-7101 (2000).</li><li>Schindelin, J., Rueden, C. T., Hiner, M. C., Eliceiri, K. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+ImageJ+ecosystem:+An+open+platform+for+biomedical+image+analysis.">The ImageJ ecosystem: An open platform for biomedical image analysis.</a> Molecular Reproduction and Development. 82, 518-529 (2015).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=R:+A+Language+and+Environment+for+Statistical+Computing.">R: A Language and Environment for Statistical Computing.</a> R Core. , (2017).</li><li>. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RStudio:+Integrated+Development+Environment+for+R.">RStudio: Integrated Development Environment for R.</a> R Team. , (2016).</li><li>. nandb R package Available from: <a target="_blank" href="https://CRAN.R-project.org/package=nandb">https://CRAN.R-project.org/package=nandb</a> (2017)</li><li>Jung, G., Wiehler, J., Zumbusch, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+photophysics+of+green+fluorescent+protein:+influence+of+the+key+amino+acids+at+positions+65,+203,+and+222.">The photophysics of green fluorescent protein: influence of the key amino acids at positions 65, 203, and 222.</a> Biophysical Journal. 88, 1932-1947 (2005).</li><li>Shaner, N. C., Steinbach, P. A., Tsien, R. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+guide+to+choosing+fluorescent+proteins.">A guide to choosing fluorescent proteins.</a> Nature Methods. 2, 905-909 (2005).</li><li>Butkevich, A. N., 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=Hydroxylated+fluorescent+dyes+for+live-cell+labeling:+synthesis,+spectra+and+super-resolution+STED.">Hydroxylated fluorescent dyes for live-cell labeling: synthesis, spectra and super-resolution STED.</a> Chemistry: A European Journal. 23, 12114-12119 (2017).</li></ol>

N & B 是一种检测 multimerization 的技术, 使用商用光扫描共聚焦显微镜配备数字探测器。与单点 FCS、FCCS 和 smFRET 相比, 该方法具有很好的吸引力, 因为它是无标定的, 亮度计算简单, 浓度独立6。然而, 在执行亮度计算9之前, 纠正漂白和长期强度波动是很重要的;如果 detrending 被忽略或不正确地执行, 由于漂白和其他工件的亮度轻微增加可能被误解为寡聚状态的变化 (如...

蛋白质-蛋白质相互作用I n-体外
传统上, 晶体学和核磁共振实验结合低温电子显微镜 (cryoEM) 是选择准确描述蛋白质三维结构的技术, 并推断其功能审议他们的高分辨率结构细节。然而, 蛋白质不是静止的结构, 可以经受各种构象变化和时间和空间的振动。这就是为什么来自晶体或 CryoEM 数据的结构信息需要辅以其他技术 (例如,分子动力学模拟和单分子技术): 蛋白质的功能与其构象有关。更改和交互, 并且此信息不存在于静态结构中。为了探索分子内动力学, 基于单分子福斯特共振能量转移 (smFRET) 的技术是非常有效的1。这些方法能够评估复杂介质中分子的不同亚群。这是非常重要的, 因为这些变化是迅速的, 并发生在获取数据 (即纳秒到第二个范围)。
通常使用两种主要方法来检测和量化这些变化: 溶液中的蛋白质和表面固定化。对于分子间相互作用的检测, 特别是配体诱导的二聚化过程, smFRET 并不总是最佳的工具。的确, 烦恼不仅依赖于距离 (≈10 nm), 而且还取决于两个偶极子 (施主和受体, χ2) 的方向和供体发射与受体的吸收光谱2的重叠, 但也许这最后的条件是少重要的是, 如果实验家可以选择正确的焦虑夫妇。smFRET 探测二聚化的一个特殊缺点来自于对感兴趣的蛋白质的标记: 对于异性 smFRET, 二聚化只能检测到 50% (即,异种焦虑只能检测到捐赠者受体和受体-捐赠者的脂肪酸, 但不是捐助者或受体-受体, 这是其他50% 的脂肪酸)。利用荧光相关光谱学 (FCS) 和衍生物 (FCCS等3) 来确定蛋白质的扩散常数和体外结合常数是另一种选择。这些方法无法完全量化二聚化, 就像在 FCS 一测量浓度和扩散, 扩散粒子的半径和扩散系数非常依赖于分子量;例如, 分子量的10倍增加只意味着扩散系数4的2.15 倍变化。在双色 FCS 或 FCCS 的情况下, 只有50% 的脂肪酸将被视为与上述相同的原因。在体外和体内检测二聚化的最实用和定量的方法是5 、数字和亮度 (N & B)6。鉴于人类的焦虑需要特定的仪器恢复的各向异性值 (即光学元件/分析仪, 以恢复平行和垂直极化), N & B 在这里提出作为一个有利的技术, 以检测蛋白质二聚化和聚合。它可以在体外和体内使用商业设置。
数字和亮度
N & B 最近被审查了7。该评价的重点是技术在活细胞中的应用。这是值得在这里重现的数学形式主义, 因为这些等式将应用于收集的数据体外。首先, 有必要定义一些术语和数学量:
<ul><li>实体是一组被绑定在一起的分子。</li>
	<li>一个实体的亮度ε是每单位时间 (每帧) 发出的光子数。</li>
	<li>
n是存在的实体数。</li>
	<li>对于图像序列中的给定像素, 是其平均强度, σ2是其强度的方差。</li>
</ul>然后, 用光子计数探测器和假设移动实体, 没有背景,
<img alt="Equation 1" src="/files/ftp_upload/58157/58157eq1.jpg"> <img alt="Equation 2" src="/files/ftp_upload/58157/58157eq2.jpg">
其中N是明显的数字, B是明显的亮度。这会导致
<img alt="Equation 3" src="/files/ftp_upload/58157/58157eq3.jpg"> <img alt="Equation 4" src="/files/ftp_upload/58157/58157eq4.jpg">
Dalal等。8表明, 与模拟设备, 你需要三更正术语: S 因素, 背景偏移, 和读数噪声σ02。然后, 再次假设移动实体,
<img alt="Equation 5" src="/files/ftp_upload/58157/58157eq5.jpg"> <img alt="Equation 6" src="/files/ftp_upload/58157/58157eq6.jpg">
给
<img alt="Equation 7" src="/files/ftp_upload/58157/58157eq7.jpg"> <img alt="Equation 8" src="/files/ftp_upload/58157/58157eq8.jpg">
请注意, 上述等式是不同的, 给在 Dalal等。8和随后的审查。7在 Dalal等, 分母中的S被省略了由于一个错字和这个错误被复制在审查。上面的等式是正确的。测量S, 偏移和σ02的指示-连同他们的意思解释-是由 Dalal等人给出的。8
亮度ε是成正比的寡聚状态的扩散实体: ε将是两倍大的脂肪酸, 因为它是对单体, 三倍大的三聚体, 因为它是为单体, 两倍大的 hexamers, 因为它是三聚体等。这样, 测量亮度ε, 你可以量化任何类型的 multimerization。
如果存在寡聚状态的混合物, 则数字和亮度无法恢复所呈现的单个寡聚状态。这是技术的局限。
Detrend 算法与 nandb 软件
对漂白的纠正的重要性以前曾被强调过9。漂白在光镜实验中不可避免地出现时间失效模式;在活细胞和体外。在文献中已经描述了许多方法, 以纠正漂白7。detrending 参数T的自动选择指数滤波技术是目前最好的方法。它集成到免费的开源软件 nandb9中。实际上, 需要用户手动选择其 detrending 参数的软件可能会导致错误的结果, 因为此参数选择可能是任意的和不正确的。自动算法检查数据并确定相应的参数, 不需要用户干预9。即使使用平滑参数的最佳选择, detrending 也有其局限性, 并且仅在小于25% 的漂白百分比下工作, 如模拟9所示。有趣的是, 当使用自动 detrending 例程时, 它的精确度是这样的, 你可以使用低亮度值 (即使是B < 1.01), 因此低强度, 仍然是精确到量化的二聚化。
漂白还导致另一个问题: photobleached 显影存在于 multimer 复杂中。这使例如,当三聚体中的三个单位中的一个是非荧光的时候, 三聚体就像一个二聚体。宾虚和米勒10显示了如何纠正这一点, 这一修正也强调在随后的审查7。nandb 软件包括此更正9。
FKBP12F36V系统
FKBP12F36V 是一种不自然 oligomerize 的蛋白质, 但 dimerize 在 AP20187 药物 (俗称 BB dimerizing 配体)11、12的加入后才知道。这使得它成为一个理想的数字和亮度测试用例: 用标记的 FKBP12F36V, 寡聚状态的加倍应该观察在加法 BB。

<table border="0" cellpadding="0" cellspacing="0" style="width:997px;" width="997">
	<tbody>
		<tr height="40">
			<td height="40" style="height:40px;width:331px;">RosettaTM (DE3) pLysS cells</td>
			<td style="width:123px;">Novagen</td>
			<td style="width:344px;">70956-3</td>
			<td style="width:200px;"></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Ampicillin</td>
			<td style="width:123px;">Sigma Aldrich</td>
			<td style="width:344px;">PubChem Substance ID 329824407</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Chloramphenicol</td>
			<td style="width:123px;">Sigma Aldrich</td>
			<td>PubChem Substance ID: 24892250</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">LB starter culture</td>
			<td style="width:123px;">QIAGEN</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">LB medium</td>
			<td style="width:123px;">QIAGEN</td>
			<td height="40" style="height:40px;width:344px;&quot;">https://www.sigmaaldrich.com/content/dam/sigma-aldrich/head/search/external-link-icon.gif</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">IPTG</td>
			<td style="width:123px;">Sigma Aldrich</td>
			<td style="width:344px;">PubChem Substance ID 329815691</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">IMAC buffer</td>
			<td>Medicago</td>
			<td>09-1010-10</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">EDTA-free protease inhibitors&#160;</td>
			<td style="width:123px;">Sigma Aldrich</td>
			<td>11873580001</td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">TALON resin</td>
			<td>Clonetech</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Nickel sepharose</td>
			<td style="width:123px;">GE Healthcare</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">S200 16/60 column</td>
			<td style="width:123px;">GE Healthcare</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="40">
			<td height="40" style="height:40px;">Glass bottom 8 well observation dish</td>
			<td>Ibidi</td>
			<td>80827</td>
			<td></td>
		</tr>
	</tbody>
</table>

calibration-free in vitro quantification of protein homo-oligomerization using commercial instrumentation and free, open source brightness analysis software

数字和亮度是一种无标定荧光涨落光谱 (FFS) 技术, 用于检测蛋白质的分子齐聚。它可以使用传统的共焦显微镜配备数字探测器。在体外使用该技术的一项协议通过一个使用案例显示, 在该用例中, 数字和亮度可以准确量化 mVenus 标记的 FKBP12F36V 在 dimerizing 药物 AP20187 添加之前和之后的寡聚状态。讨论了使用正确的显微采集参数和正确的数据预处理和分析方法的重要性。尤其强调了选择漂白校正的重要性。这种廉价的方法可以用来研究蛋白质与蛋白质之间的相互作用, 在许多生物环境。

Detrending 和单体亮度
获取数据后, 可以开始亮度计算, 以确定解决方案中感兴趣的蛋白质的寡聚状态。即使漂白在溶液中的作用可能不会像体内那样剧烈, 但强度痕迹仍可能没有固定的平均值, 可能是由于与荧光有关的物理效应,18但背后的原因是不完全理解的。这在计算亮度时有影响;当试图?...

Watch this Scientific Journal Video about Calibration-free In Vitro Quantification of Protein Homo-oligomerization Using Commercial Instrumentation and Free, Open Source Brightness Analysis Software a

Calibration-free In Vitro Quantification of Protein Homo-oligomerization Using Commercial Instrumentation and Free, Open Source Brightness Analysis Software

利用商用仪器和自由开源亮度分析软件对蛋白质的聚齐聚进行无标定的体外定量化

本协议描述了一种无标定的方法, 用于在荧光涨落光谱的基础上, 利用商用光扫描显微镜定量测定蛋白质的体外齐聚。给出了正确的采集设置和分析方法。

Number and brightness is a calibration-free fluorescence fluctuation spectroscopy (FFS) technique for detecting protein homo-oligomerization. It can be ...

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Calibration-free In Vitro Quantification of Protein Homo-oligomerization Using Commercial Instrumentation and Free, Open Source Brightness Analysis Software

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使用脚手架脂质体重构脂质近端蛋白质 - 蛋白质相互作用<em&gt;体外</em

Studies of integral membrane proteins in vitro are frequently complicated by the presence of a hydrophobic transmembrane domain. Further complicating ...

科研

生物化学

Biochemical and Structural Characterization of the Carbohydrate Transport Substrate-binding-protein SP0092

碳水化合物转运基质结合蛋白 SP0092 的生化和结构表征

Development of new antimicrobials and vaccines for Streptococcus pneumoniae (pneumococcus) are necessary to halt the rapid rise in multiple resistant ...

Quantification of Hypopigmentation Activity In Vitro

低聚活性的体外定量

This study presents laboratory methods for the quantification of hypopigmentation activity in vitro. Melanin, the major pigment in melanocytes, is ...

Analysis of Protein Folding, Transport, and Degradation in Living Cells by Radioactive Pulse Chase

放射性脉冲追逐对活细胞蛋白质折叠、迁移和降解的分析

Radioactive pulse-chase labeling is a powerful tool for studying the conformational maturation, the transport to their functional cellular location, and ...

Using the Open-Source MALDI TOF-MS IDBac Pipeline for Analysis of Microbial Protein and Specialized Metabolite Data

使用开源 MALDI TOF-MS IDBac 管道分析微生物蛋白和专用代谢物数据

In order to visualize the relationship between bacterial phylogeny and specialized metabolite production of bacterial colonies growing on nutrient agar, ...

Quantification of Protein Interaction Network Dynamics using Multiplexed Co-Immunoprecipitation

使用多路复用共免疫沉淀对蛋白质相互作用网络动力学进行量化

Dynamic protein-protein interactions control cellular behavior, from motility to DNA replication to signal transduction. However, monitoring dynamic ...

Absolute Quantification of Cell-Free Protein Synthesis Metabolism by Reversed-Phase Liquid Chromatography-Mass Spectrometry

通过反相液相-质谱法对无细胞蛋白合成代谢的绝对定量

Cell-free protein synthesis (CFPS) is an emerging technology in systems and synthetic biology for the in vitro production of proteins. However, if CFPS is ...

Quantification of Humic and Fulvic Acids in Humate Ores, DOC, Humified Materials and Humic Substance-Containing Commercial Products

腐植酸盐矿石、DOC、腐殖化材料和含腐殖质的商业产品中腐殖酸和黄腐酸的定量

The purpose of this method is to provide an accurate and precise concentration of humic (HA) and/or fulvic acids (FA) in soft coals, humic ores and ...

Laser-free Hydroxyl Radical Protein Footprinting to Perform Higher Order Structural Analysis of Proteins

无激光羟基基蛋白足迹，对蛋白质进行更高级的结构分析

Hydroxyl Radical Protein Footprinting (HRPF) is an emerging and promising higher order structural analysis technique that provides information on changes ...

Making Precise and Accurate Single-Molecule FRET Measurements using the Open-Source smfBox

使用开源 smfBox 进行精确和准确的单分子 FRET 测量

The smfBox is a recently developed cost-effective, open-source instrument for single-molecule F&#246;rster Resonance Energy Transfer (smFRET), which makes ...