Name: synthesis of an in vivo mri-detectable apoptosis probe
Uploaded: 2012-07-31T12:00:00
Description: Watch this Scientific Journal Video about Synthesis of an In vivo MRI-detectable Apoptosis Probe at JoVE.com

国家机构的健康，NHLBI（市，PY，PS）。 美国心脏协会（巨浪）。 斯坦福大学，VPUE格兰特（巨浪）。

改编自以前的研究1。 1。 Annexin V的共轭超顺磁性氧化铁<ol><li>氧化0.15 M高碘酸钠（NAIO 4）（4:1重量：重量比）为1小时在黑暗中的解决方案与超顺磁性氧化铁粒子（纳米技术公司，海洋5毫克/毫升）在20°C 2。 </li><li>在20°C孵育氧化12小时的超顺磁性氧化铁，用纯化的的ANX蛋白（比例为1:1，重量：重量）0.15 M四硼酸钠（钠2 B 4 O 7 10H 2） <...

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	<li>Dash, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+molecular+MRI+probe+to+detect+treatment+of+cardiac+apoptosis+in+vivo.">A molecular MRI probe to detect treatment of cardiac apoptosis in vivo.</a> Magn. Reson. Med. , (2011).</li><li>Khym, J. X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+reaction+of+methylamine+with+periodate-oxidized+adenosine+5'-phosphate.">The reaction of methylamine with periodate-oxidized adenosine 5'-phosphate.</a> Biochemistry. 2, 344-350 (1963).</li><li>Liu, Y. Y., Yu, Y., Zhang, G. B., Tang, M. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Non-invasive+detection+of+apoptosis+using+magnetic+resonance+imaging+and+a+targeted+contrast+agent.">Non-invasive detection of apoptosis using magnetic resonance imaging and a targeted contrast agent.</a> Nat. Med. 7, 1241-1244 (2001).</li><li>van Tilborg, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Internalization+of+annexin+A5-functionalized+iron+oxide+particles+by+apoptotic+Jurkat+cells.">Internalization of annexin A5-functionalized iron oxide particles by apoptotic Jurkat cells.</a> Contrast Media Mol. Imaging. 4, 24-32 (2009).</li><li>Sosnovik, D. 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连接超顺磁性氧化铁Annexin V的共轭方法利用膜联蛋白的侧链胺组和超顺磁性氧化铁纳米粒子的羧基基团。通过特定的氧化还原步骤，这些化合物的共价键可以实现，产生的功能化纳米粒子可以被孤立。此法可推广到其他蛋白质与纳米粒子的利益。 最关键的步骤是氧化还原条件，在适当的温度和温和搅拌。产量低，可能会导致不完全氧化或减少，导致无残留Annexin V的，在过滤?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录＃ </td></tr><tr><td>超顺磁性氧化铁</td><td>海洋纳米技术公司</td><td> ICK-40-005 </td></tr><tr><td>阿霉素</td><td>西格玛</td><td> D1515-10MG </td></tr><tr><td> SuperMag分离</td><td>海洋纳米技术公司</td><td> N / A </td></tr><tr><td>动态光散射纳米激光粒度仪机</td><td>马尔文公司</td><td> N / A </td></tr></tbody></table>

synthesis of an in vivo mri-detectable apoptosis probe

细胞凋亡是许多疾病的显着特征，这一程序性细胞死亡之前，通常会发生疾病的临床表现是显而易见的。在其最早的，可逆的阶段检测细胞凋亡的一种手段，将承担临床前的&#39;窗口&#39;，预防或治疗措施，在此期间，可以采取保护心脏的永久性损伤。这里我们提出一个简单和可靠的方法，共轭人Annexin V的（ANX），如饥似渴地结合到细胞中最早的，可逆的凋亡阶段，超顺磁性氧化铁（SPIO）纳米粒子，作为检测的MRI造影剂。共轭方法开始的超顺磁性氧化铁纳米粒子，氧化成羧基的超顺磁性氧化铁多糖壳的氧化。纯化蛋白ANX，然后添加一个硼酸钠溶液，以方便与超顺磁性氧化铁在ANX共价相互作用，减少缓冲区的设置。一个与钠borohydrid最后削减步骤e是执行完成减少，然后淬火反应。未结合ANX从离心过滤组合。动态光散射（DLS）的大小和ANX-超顺磁性氧化铁产品纯度验证。这种方法并不需要，或修改，超顺磁性氧化铁壳多糖，反对交联氧化铁粒子共轭方法或生物素标记的纳米粒子。因此，这种方法代表了一个简单，可靠的方法，可以延长至共轭利益的其他蛋白质。

Watch this Scientific Journal Video about Synthesis of an In vivo MRI-detectable Apoptosis Probe at JoVE.com

Synthesis of an In vivo MRI-detectable Apoptosis Probe

细胞凋亡的早期检测可以识别处于危险的细胞群，在多种疾病。在这里，我们展示了一个方法来链接早期凋亡检测蛋白Annexin V的一个MRI检测的氧化铁纳米粒子（超顺磁性氧化铁）。此方法可扩展到其他感兴趣的蛋白质生成MRI检测的分子成像探针。

合成的<em&gt;在体内</em&gt; MRI检测细胞凋亡的探讨

Cellular apoptosis is a prominent feature of many diseases, and this programmed cell death typically occurs before clinical manifestations of disease are ...

Research

JoVE Journal

Biology

Synthesis of an In vivo MRI-detectable Apoptosis Probe

Homemade Site Directed Mutagenesis of Whole Plasmids

自制的网站定向诱变整个质粒

Site directed mutagenesis of whole plasmids is a simple way to create slightly different variations of an original plasmid. With this method the cloned ...

科研

生物学

Photoconversion of Purified Fluorescent Proteins and Dual-probe Optical Highlighting in Live Cells

在活细胞荧光蛋白的双探头光学突出纯净的光转化的

Photoconvertible fluorescent proteins (pc-FPs) are a class of fluorescent proteins with "optical highlighter" capability, meaning that the color of ...

In vivo Imaging and Therapeutic Treatments in an Orthotopic Mouse Model of Ovarian Cancer

In vivo Imaging and Therapeutic Treatments in an Orthotopic Mouse Model of Ovarian Cancer

活体成像和治疗卵巢癌的原位小鼠模型

Human cancer and response to therapy is better represented in orthotopic animal models.  This paper describes the development of an orthotopic mouse model ...

Monitoring Dynamic Changes In Mitochondrial Calcium Levels During Apoptosis Using A Genetically Encoded Calcium Sensor

使用一种基因编码的钙传感器来监测细胞凋亡过程中线粒体钙水平的动态变化

Dynamic changes in intracellular calcium concentration in response to various stimuli regulates many cellular processes such as proliferation, ...

Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c

Activation of Apoptosis by Cytoplasmic Microinjection of Cytochrome c

激活细胞凋亡的细胞色素的细胞质显微注射 C</em

Apoptosis, or programmed cell death, is a conserved and highly regulated pathway by which cells die1.  Apoptosis can be triggered when cells encounter a ...

The Tomato/GFP-FLP/FRT Method for Live Imaging of Mosaic Adult Drosophila Photoreceptor Cells

The Tomato/GFP-FLP/FRT Method for Live Imaging of Mosaic Adult Drosophila Photoreceptor Cells

番茄/ GFP-FLP / FRT方法马赛克成人的实时成像<em&gt;果蝇</em&gt;感光细胞

The Drosophila eye is widely used as a model  for studies of development and neuronal degeneration. With the powerful mitotic  recombination technique, ...

Analysis of Apoptosis in Zebrafish Embryos by Whole-mount Immunofluorescence to Detect Activated Caspase 3

Whole-mount immunofluorescence to detect activated Caspase 3 (Casp3 assay) is useful to identify cells undergoing either intrinsic or extrinsic apoptosis ...

An in vivo Crosslinking Approach to Isolate Protein Complexes From Drosophila Embryos

An in vivo Crosslinking Approach to Isolate Protein Complexes From Drosophila Embryos

一个在体内交联方法分离蛋白复合物从果蝇胚胎

Many cellular processes are controlled by multisubunit protein complexes. Frequently these complexes form transiently and require native environment to ...

An Ex vivo Culture System to Study Thyroid Development

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一个<em&gt;体外</em&gt;企业文化系统来研究甲状腺发展

The thyroid is a bilobated endocrine gland localized at the base of the neck, producing the thyroid hormones T3, T4, and calcitonin. T3 and T4 are ...

Strategies for Tracking Anastasis, A Cell Survival Phenomenon that Reverses Apoptosis

策略跟踪Anastasis，A细胞存活的现象，细胞凋亡反转

Anastasis (Greek for &#8220;rising to life&#8221;) refers to the recovery of dying cells. Before these cells recover, they have passed through important ...