Name: direct measurement of kdm1a target engagement using chemoprobe-based immunoassays
Uploaded: 2019-06-13T13:00:00
Description: Watch this Scientific Journal Video about Direct Measurement of KDM1A Target Engagement Using Chemoprobe-based Immunoassays at JoVE.com

这项研究由奥里松基因组学资助。S.A.,霍夫曼-拉罗氏,部分支持CIIP-20152001和RETOS合作项目RTC-2015-3332-1。

根据西班牙立法(第1716/2011年《生物资源研究法》)并经当地道德委员会批准,从圣保生物库研究所获得血液样本。动物组织研究是根据动物实验伦理委员会制定的关于护理和使用实验室动物的机构准则(欧洲共同体理事会第86/609/EEC号指令)进行的。普拉-多氯联苯。
1. 制备用于检测的生物样本。
注意:本协议涉及操纵可能受职业安全与健康管理局 (OSHA) 血?...

<ol>
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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DNA+Hypomethylation+Signature+Predicts+Antitumor+Activity+of+LSD1+Inhibitors+in+SCLC.">DNA Hypomethylation Signature Predicts Antitumor Activity of LSD1 Inhibitors in SCLC.</a> Cancer Cell. 28 (1), 57-69 (2015).</li><li>Rivers, 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=RN-1,+a+potent+and+selective+lysine-specific+demethylase+1+inhibitor,+increases+&#947;-globin+expression,+F+reticulocytes,+and+F+cells+in+a+sickle+cell+disease+mouse+model.">RN-1, a potent and selective lysine-specific demethylase 1 inhibitor, increases &#947;-globin expression, F reticulocytes, and F cells in a sickle cell disease mouse model.</a> Experimental Hematology. 43 (7), 546-553 (2015).</li><li>Rivers, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oral+administration+of+the+LSD1+inhibitor+ORY-3001+increases+fetal+hemoglobin+in+sickle+cell+mice+and+baboons.">Oral administration of the LSD1 inhibitor ORY-3001 increases fetal hemoglobin in sickle cell mice and baboons.</a> Experimental Hematology. 67, 60-64 (2018).</li><li>Buesa, 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=The+dual+LSD1/MAO-B+inhibitor+ORY-2001+prevents+the+development+of+the+memory+deficit+in+samp8+mice+through+induction+of+neuronal+plasticity+and+reduction+of+neuroinflammation.">The dual LSD1/MAO-B inhibitor ORY-2001 prevents the development of the memory deficit in samp8 mice through induction of neuronal plasticity and reduction of neuroinflammation.</a> Alzheimer&#8217;s &amp; Dementia. 11 (7), P905 (2015).</li><li>Schmidt, D. 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T., Cockerill, M. J., Hitchin, J. R., Wiseman, D. H., Somervaille, T. 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此处介绍的协议是使用基于新型 KDM1A 化学探针捕获的 ELISA 直接测量 KDM1A 目标参与度的。该方法已对培养的人类细胞系和来自人类、大鼠和小鼠和巨蜥(包括PBMC、肺、大脑、皮肤、肿瘤)的体外样本进行了验证,但可随时应用于KDM1A抗体靶向表位和催化的其他物种中心是保存的。由于OG-881是一种基于活性的化学探针,样品质量很重要,因此,特别是在协议的初始步骤中,应进行适当的操作和保护,以确保KDM1A...

莱辛特异性脱甲基酶1(KDM1A)1是一种参与控制基因转录的脱甲基酶。这种蛋白质已成为肿瘤学的候选药理靶点2;包括急性骨髓性白血病3(AML),骨髓增生综合征(MDS)4,骨髓纤维化(MF)5,6,小细胞肺癌(SCLC)7;在病变细胞病(SCD)8,9,和在中枢神经系统疾病,包括阿尔茨海默氏病(AD),多发性硬化症(MS);并在侵略10。
临床开发中的大多数KDM1A抑制化合物是环丙胺衍生物,通过共价结合其片苷二核苷酸(FAD)联因子11来抑制蛋白质。KDM1A的抑制诱导基因表达变化,但这些变化在组织、细胞类型或疾病病例之间变化很大。抑制KDM1A也改变组蛋白标记12,但这些变化一般在基因组的特定部位本地产生,并再次是高度组织和细胞特异性。
该协议旨在直接测量KDM1A目标在生物样品中的参与度,并针对环丙胺衍生抑制剂的使用进行了优化。该测定基于ELISA技术,并并行分析从生物样品中提取的天然蛋白质提取物中的KDM1A(非约束抑制剂)。作为第一步,生物样品在存在生物异化KDM1A选择性化学探针OG-88113,14,来自选择性KDM1A抑制剂ORY-1001(iadademstat),一种临床KDM1A的强效抑制剂的存在下进行。肿瘤病的治疗发展。化学探针具有 IC50,用于 KDM1A 的 120 nM,并包括与生物素化聚乙烯乙二醇 (PEG) 尾部相关的 FAD 结合功能。化学探针专门绑定到游免费的KDM1A,但不与样品中的抑制剂结合的KDM1A结合。在化学探针结合后,在具有链球菌素涂层表面的微子板上捕获含有复合物的KDM1A,以确定游脱KDM1A,或在涂有单克隆抗KDM1A捕获抗体的板上捕获,以确定总KDM1A。洗涤后,两个板均用抗KDM1A检测抗体孵育,再次洗涤,并用二级HRP结合的驴抗兔IgG抗体孵育,使用发光基板进行检测,并通过测量相对光照仪 (RLU) 中的光度计 (图 1)。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/59390/59390fig1.jpg"> 图 1.ELISA酶与化学探针免疫吸收测定的架构为KDM1A靶点:A)使用三明治ELISA和B确定总KDM1A(使用化学探针ELISA)的游出KDM1A。<a href="https://www.jove.com/files/ftp_upload/59390/59390fig1large.jpg" target="_blank">请点击此处查看此图的较大版本。</a>
两个 ELISA 板中都包含一条标准曲线,用于验证每个测定的线性度。然后,将确定每个样品中的KDM1A目标参与度作为预剂量或车辆处理样品的相对值计算。

<table>
	<tbody>
		<tr>
			<td>0,05% Trypsin-EDTA (1X)</td>
			<td>Thermo Scientific</td>
			<td>#25300-062</td>
			<td></td>
		</tr>
		<tr>
			<td>10 X Protease Inhibitor Tablets</td>
			<td>Roche</td>
			<td>#11836153001</td>
			<td></td>
		</tr>
		<tr>
			<td>96 deep well storage block</td>
			<td>VWR</td>
			<td>#734-1679</td>
			<td></td>
		</tr>
		<tr>
			<td>96 well ELISA plates</td>
			<td>Nunc</td>
			<td>#436110</td>
			<td></td>
		</tr>
		<tr>
			<td>Adhesive black Film</td>
			<td>Perkin Elmer</td>
			<td>#6050173</td>
			<td></td>
		</tr>
		<tr>
			<td>Adhesive transparent Film</td>
			<td>VWR</td>
			<td>#60941-062</td>
			<td></td>
		</tr>
		<tr>
			<td>Biotinylated KDM1A probe OG-881</td>
			<td>Oryzon Genomics S.A.</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin</td>
			<td>Sigma</td>
			<td># 3117057001</td>
			<td></td>
		</tr>
		<tr>
			<td>Bovine Serum Albumin Standard</td>
			<td>Thermo Scientific</td>
			<td>#23208)</td>
			<td></td>
		</tr>
		<tr>
			<td>Bradford Protein Assay</td>
			<td>BioRad</td>
			<td>#500-0001</td>
			<td></td>
		</tr>
		<tr>
			<td>Cell lysis buffer 10X</td>
			<td>Cell Signaling</td>
			<td>#9803</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge for 96- well plates</td>
			<td>Hettich</td>
			<td>Rotina 420R</td>
			<td></td>
		</tr>
		<tr>
			<td>Flask</td>
			<td>Thermo Scientific</td>
			<td>#156499</td>
			<td></td>
		</tr>
		<tr>
			<td>Full length, enzymatically active human Recombinant LSD1 / KDM1A</td>
			<td>Active Motif</td>
			<td>#31426</td>
			<td></td>
		</tr>
		<tr>
			<td>Graphpad Prism 5 Project</td>
			<td>GraphPad Software</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>Luminol-Enhacer and Peroxide Solution (Chemiluminescent Substrate)</td>
			<td>Thermo Scientific</td>
			<td>#37074</td>
			<td></td>
		</tr>
		<tr>
			<td>Micro Centrifuge</td>
			<td>Eppendorf</td>
			<td>5415 R</td>
			<td></td>
		</tr>
		<tr>
			<td>Microplate reader Infinite 200-Tecan</td>
			<td>Tecan</td>
			<td>Infinite 200</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouse monoclonal capture antibody Anti-KDM1A (N-terminal epitope)</td>
			<td>Abcam</td>
			<td>#ab53269</td>
			<td></td>
		</tr>
		<tr>
			<td>Needle G18 gauge blunt</td>
			<td>BD</td>
			<td>#303129</td>
			<td></td>
		</tr>
		<tr>
			<td>ORY-1001 (iadademstat)</td>
			<td>Oryzon Genomics S.A.</td>
			<td>NA</td>
			<td></td>
		</tr>
		<tr>
			<td>PBMC separation tubes 10 ml</td>
			<td>Greiner bio-one</td>
			<td>#163288</td>
			<td></td>
		</tr>
		<tr>
			<td>PBMC separation tubes 50 ml</td>
			<td>Greiner bio-one</td>
			<td>#227288</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS 1x</td>
			<td>Sigma</td>
			<td>#D8537</td>
			<td></td>
		</tr>
		<tr>
			<td>Plate shaker</td>
			<td>Heidolph Instruments</td>
			<td>Rotamax 120</td>
			<td></td>
		</tr>
		<tr>
			<td>Polysorbate 20</td>
			<td>Sigma</td>
			<td>#P7949</td>
			<td></td>
		</tr>
		<tr>
			<td>Rabbit monoclonal detection antibody Anti-KDM1A (C-terminal epitope)</td>
			<td>Cell Signaling</td>
			<td>#672184BF-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Secondary antibody Peroxidase-conjugated Donkey Anti-rabbit IgG</td>
			<td>Thermo Scientific</td>
			<td>#31458</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrophotometer cuvette 1.5</td>
			<td>Deltalab</td>
			<td>#302100</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectrophotometer for cuvette</td>
			<td>GE Healthcare</td>
			<td>GeneQuant 1300</td>
			<td></td>
		</tr>
		<tr>
			<td>Streptavidin</td>
			<td>Promega</td>
			<td>#Z704A</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe</td>
			<td>BD</td>
			<td>#303172</td>
			<td></td>
		</tr>
		<tr>
			<td>Type 1 ultrapure water</td>
			<td>Millipore</td>
			<td>Milli-Q Advantage A10</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultrasonic cleaner</td>
			<td>VWR</td>
			<td>USC200T</td>
			<td></td>
		</tr>
	</tbody>
</table>

direct measurement of kdm1a target engagement using chemoprobe-based immunoassays

对靶向参与的评估,即药物与其设计的蛋白质相互作用,是解释药物开发或基础研究项目中任何化合物的生物活性的基本要求。在表观遗传学中,目标参与度通常通过分析代理标记而不是测量化合物与目标的结合来评估。已分析的下游生物读出包括组蛋白标记调制或基因表达变化。KDM1A是一种以源脱甲基酶,从单甲基和二甲基化H3K4中去除甲基组,这是一种与基因表达沉默相关的修饰。代理标记的调制取决于被调查细胞的细胞类型和功能,这使得解释和交叉案例比较相当困难。为了规避这些问题,提出了一个通用的协议,以评估直接KDM1A目标参与的剂量效应和动态。所述测定利用KDM1A化学探针捕获和量化无抑制酶,可广泛应用于细胞或组织样本,无需基因改造,具有极好的检测窗口,可用于基础研究临床样本分析。

总和游免费 KDM1A 测定的线性度。
标准系列是编写步骤5.3.2.的,使用0至2500皮克的全长人体重组KDM1A酶。评估了总和自由 rKDM1A 的 RLU 值以验证线性度(图 2A和2B)。数据表示为 3 个实验的平均值,3 个技术复制 (n) = SD。在3个独立志愿者的血液中检测出人类PBMC中总和游离KDM1A的RLU值被叠加?...

Watch this Scientific Journal Video about Direct Measurement of KDM1A Target Engagement Using Chemoprobe-based Immunoassays at JoVE.com

Direct Measurement of KDM1A Target Engagement Using Chemoprobe-based Immunoassays

使用基于化学探针的免疫测定直接测量KDM1A目标参与

在这里,我们提出一个协议,以测量KDM1A靶向在使用KDM1A抑制剂处理的人或动物细胞、组织或血液样本中的参与度。该协议采用自由KDM1A酶的化学探针标记,并使用基于化学探针的免疫测定直接定量目标职业,可用于临床前和临床研究。

The assessment of the target engagement, defined as the interaction of a drug with the protein it was designed for, is a basic requirement for the ...

评估目标参与度，即药物与它所设计的蛋白质的相互作用，是解释药物开发或基础研究项目中任何化合物的生物活性的基本要求。该技术的主要优点是，它允许直接测量KDM1A目标接触的剂量效应和动力学，而不是测量下游效应，如组蛋白标记和表达。该技术可应用于基础研究和临床试验，在肿瘤学中 CNS 或其他受 KDM1A 抑制剂治疗的疾病中，用于研究药理学。该技术基于一种化疗开发，用于研究 ORY-1001，iadademstat，但可以真正用于其他KDM1A抑制剂，该策略可应用于其他靶点。该程序将由我实验室的PK/PD发现平台的操作经理Raquel Ruiz演示。在用 ORY-1001 进行细胞治疗期间，化合物与细胞核中的 KDM1A 蛋白结合。为了开始此过程，从冰箱中取出 10 微升一次使用 20 毫摩尔生物基化探针 OG-881 库存溶液，让它在室温下预热 10 分钟。使用带滤芯的微管，连续稀释库存溶液，以准备两个微摩尔工作溶液。接下来，在微离心管中溶解一片蛋白酶抑制剂在一毫升PBS中。对于每毫升所需的1x细胞解液缓冲液与化疗，混合100微升商业获得的10x细胞解液缓冲液，150微升的10x蛋白酶抑制剂，12.5微升的两微摩尔OG-881工作溶液和737.5微升1型双蒸馏水。细胞在1x解液缓冲液中，在与任何自由KDM1A蛋白结合的化疗存在的情况下。要从组织中准备，请使用在干冰上冷藏的砂浆和害虫来粉碎一立方厘米的冷冻组织并均质化。将组织分成一次使用小瓶，确保将大约40毫克的组织粉末转移到每个小瓶中，同时避免在所有的时间解冻。将样品存放在 80 摄氏度，直到准备好处理。为了从细胞颗粒中准备，在含有25纳米摩尔OG-881的1x细胞解液缓冲液的200微升中重新填充约1000万个细胞的颗粒。每个样品都短暂旋转，并把它们放在冰上五分钟。使用声波器，以 45 千赫的三个脉冲对样品进行声波化，每次持续 20 秒。将样品在冰上放置20秒的脉冲之间。将样品放在冰上再加五分钟。然后，在4摄氏度的离心机中短暂旋转，将样品以14000倍G离心10分钟。使用一毫升微管将每个上一液转移到单独的 1.5 毫升微离心管中。加工前将管子放在冰上两个小时。然后，使用文本协议中概述的布拉德福德测定来量化原生蛋白质。总板涂有抗KDM1A抗体。自由板涂有链球菌素。然后将细胞酸盐添加到两个板中，直接或通过化疗捕获 KDM1A 复合物。用抗KDM1A检测抗体和与马萝卜过氧化物酶耦合的二级抗体清洗和孵育板。最后，添加发光基板并生成信号。对于总 KDM1A ELISA，在PBS中准备10毫升KDM1A捕获抗体，最终浓度为每毫升2微克。将 100 微升转移到板的每个井中。对于免费的KDM1A ELISA，在PBS中准备10毫升的链球菌素，每盘浓度为每毫升10微克。将 100 微升转移到板的每个井中。然后，顶部用胶粘膜密封每个板，并在摄氏四度下孵育过夜。第二天，从冰箱中取出盘子，让它们在室温下平衡约45分钟。用洗涤缓冲液清洗每个盘子三次，确保每次洗涤步骤后敲击纸巾上的盘子，以去除任何残留溶液。在两个板的每个孔中添加 200 微升的阻塞缓冲液。顶部用胶膜密封板，并在室温下孵育两小时。同时，用PBS稀释先前获得的原生蛋白质提取物，以适当的浓度，并布置板，以准备一个标准曲线使用人类rKDM1A，如文本协议中概述。两小时后孵育完成，丢弃堵塞缓冲液，用洗涤缓冲液清洗板，确保每次洗涤步骤后敲击纸巾上的板，以去除任何残留溶液。在文本协议表2中的板材分布后，将适当的稀释样品转移到冷藏的96深井储存块中。保持这个块在冰上，同时将每井100微升移入总和自由的ELISA板，按照文本协议表三中的板分布。在室温下孵育一小时，然后丢弃样品，用洗涤缓冲液清洗盘子五次。以每毫升0.125微克的浓度在阻断缓冲液中准备20毫升兔子抗KDM1A检测抗体。在两个板的每个孔中加入100微升的检测抗体溶液，除了与阴性对照相对应的孔。顶部用胶膜密封板，并在室温下孵育一小时。在此之后，丢弃检测抗体溶液，用洗涤缓冲液清洗板六次。准备25毫升的二次山羊抗兔抗体HRP，以稀释1至5000的阻尼缓冲液。在微刺激板的每个孔中加入100微升的二次抗体溶液，并在室温下孵育一小时。在此孵化结束前 30 分钟，在柔和的光线条件下，将发光醇增强剂和过氧化物溶液的等件混合在琥珀色瓶中。将混合物留在室温下。当一小时的孵育完成时，丢弃二次抗体溶液，用洗涤缓冲液洗盘子六次。接下来，将100微升的发光灯工作溶液移至板材的每个孔中，确保移液速度非常慢，避免气泡形成。使用定时器控制添加溶液和板的发光测量之间的时间，并保持此时间恒定，以实现良好的间分析可重复性。顶部用胶粘膜密封板，离心机在 500 倍 G 和室温下 45 秒，以消除任何剩余的气泡。以 100 rpm 的速度将板摇床上的板孵育一分钟。然后，取出胶粘膜，将板插入微孔板读卡器中，并留3分钟，将温度稳定在25摄氏度。读取每个 ELISA 板测定的相对发光单位。从原始数据电子表格文件中保存并复制原始 RLU 值，以进行进一步分析。在这项研究中，一种基于ELISA的新型KDM1A化疗捕获用于直接测量KDM1A靶点在细胞和组织样本中的参与度。评估总和自由 rKDM1A 的 RLU 值以验证线性度。然后，从三个独立志愿者检测到的人类 PBMC 中检测到的总量和免费 KDM1A 的 RLU 值将叠加在标准曲线上。AML细胞按照提供者的建议进行培养，并处理不同浓度的车辆或 ORY-1001。原生蛋白质提取物在存在25毫摩尔OG-881化疗和0.5微克总蛋白用于执行目标参与分析的情况下获得。然后确定总计和免费 KDM1A，并计算 ORY-1001 与 KDM1A 的目标参与率相对于车辆。剂量响应KDM1A目标参与PBMC和肺部治疗大鼠与 ORY-1001口服加瓦奇，计算相对车辆组显示在这里。用25纳米摩尔OY-1001从车辆处理过的动物中提取的肺蛋白提取物进行外活孵育，产生全TE，但并没有进一步增加被治疗的大鼠的TE，每公斤 ORY-1001的30微克，证实KDM1A在体内已经完全抑制。该协议通过测量免费和总 KDM1A 来评估 KDM1A 目标参与度。请记住，它需要本地蛋白质提取。这种技术可用于不同物种的样品，以评估特定的抑制剂，也可以筛选新的抑制剂。本研究中使用的化疗也可以应用于化疗学研究KDM1A相互作用。在处理生物样品和生物活性化合物（如KDM1A抑制剂）时，请时刻记住安全工作并尊重预防措施。

Research

JoVE Journal

Biochemistry

Covalent Binding of Antibodies to Cellulose Paper Discs and Their Applications in Naked-eye Colorimetric Immunoassays

共价结合的抗体，以纤维素纸光盘及其裸眼比色法免疫分析应用

This report presents two methods for the covalent immobilization of capture antibodies on cellulose filter paper grade No. 1 (medium-flow filter paper) ...

科研

生物化学

Measurement of Basal and Forskolin-stimulated Lipolysis in Inguinal Adipose Fat Pads

测量基底和福斯柯林刺激的脂肪分解在腹股沟脂肪脂肪垫

Lipolysis is a process by which the lipid stored as triglycerides in adipose tissues are hydrolyzed into glycerol and fatty acids. This article describes ...

Utilizing a Comprehensive Immunoprecipitation Enrichment System to Identify an Endogenous Post-translational Modification Profile for Target Proteins

利用综合沉淀富集系统识别目标蛋白的内源性后向转化剖面

It is now well-appreciated that post-translational modifications (PTMs) play an integral role in regulating a protein&#39;s structure and function, which ...

Simultaneous Measurement of Superoxide/Hydrogen Peroxide and NADH Production by Flavin-containing Mitochondrial Dehydrogenases

含黄素线粒体脱氢酶同时测定超氧化物/过氧化氢和 NADH 生产

It has been reported that mitochondria can contain up to 12 enzymatic sources of reactive oxygen species (ROS). A majority of these sites include ...

Measurement of Mitochondrial Oxygen Consumption in Permeabilized Fibers of Drosophila Using Minimal Amounts of Tissue

用最小组织量测定果蝇透纤维中线粒体耗氧量

The fruit fly, Drosophila melanogaster, represents an emerging model for the study of metabolism. Indeed, drosophila have structures homologous to human ...

A Direct Force Probe for Measuring Mechanical Integration Between the Nucleus and the Cytoskeleton

测量细胞核与骨架间机械一体化的直接力探针

The mechanical properties of the nucleus determine its response to mechanical forces generated in cells. Because the nucleus is molecularly continuous ...

Using High Content Imaging to Quantify Target Engagement in Adherent Cells

利用高含量成像对增强细胞中的目标参与进行量化

Quantitating the interaction of small molecules with their intended protein target is critical for drug development, target validation and chemical probe ...

High Sensitivity Measurement of Transcription Factor-DNA Binding Affinities by Competitive Titration Using Fluorescence Microscopy

荧光显微镜竞争滴定法测定转录机-dna 结合感的高灵敏度

Accurate quantification of transcription factor (TF)-DNA interactions is essential for understanding the regulation of gene expression. Since existing ...

Cortisol Measurement in Koala (Phascolarctos cinereus) Fur

Cortisol Measurement in Koala Phascolarctos cinereus Fur

科蒂醇测量在考拉 （菲斯科拉托斯西尼鲁斯） 毛皮

Optimal methods of hormone extraction used to measure stress in animals across sample types are not always the same. Australia&#39;s iconic marsupial ...

A Semi-Quantitative Drug Affinity Responsive Target Stability (DARTS) assay for studying Rapamycin/mTOR interaction

A Semi-Quantitative Drug Affinity Responsive Target Stability DARTS assay for studying Rapamycin/mTOR interaction

用于研究拉帕霉素/mTOR相互作用的半定量药物亲和反应目标稳定性 （DARTS） 测定

Drug Affinity Responsive Target Stability (DARTS) is a robust method for detection of novel small molecule protein targets. It can be used to verify known ...