Name: batch immunostaining for large-scale protein detection in the whole monkey brain
Uploaded: 2009-07-27T17:00:00
Description: Watch this Scientific Journal Video about Batch Immunostaining for Large-Scale Protein Detection in the Whole Monkey Brain at JoVE.com

<p class="jove_content">弗兰克欧文，罗伯塔Palmour和行为科学基金会在位于西印度群岛，圣基茨，为他们继续支持我们的灵长类动物的工作的实验室工作人员，我们非常感谢。这项工作是由来自加拿大脆性X研究基金会（FXRFC）深圳和的奖学金 - 由加拿大卫生研究院（AC）和加拿大（MP）的国家工程研究理事会研究院的运营赠款支持。 </p>

<p class="jove_content">免疫组织化学是最广泛使用的技术在各种实验动物模型的大脑蛋白的表达特征之一。这是比较容易进行系统的免疫程序，对大脑的大小相似的老鼠与其他常见的实验模型的大脑。然而，有没有发表的作品，以我们的知识，提供了一个综合考虑如何开展在整个猴脑等免疫检测程序。以下是详细描述了如何准备进行大规模的各种目标蛋白的免疫组织化学检测整个猴脑。这项工作已经成为商业和学术的?...

<ol>
	<li>Olfert, E. D., Cross, B. M., McWilliam, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> <em style='font-style: italic'>Guide to the Care and Use of Experimental Animals</em>. , (1993).</li><li>Devys, D., Lutz, Y., Rouyer, N., Bellocq, J. P., Mandel, 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=The+FMR-1+protein+is+cytoplasmic,+most+abundant+in+neurons+and+appears+normal+in+carriers+of+a+fragile+X+premutation.">The FMR-1 protein is cytoplasmic, most abundant in neurons and appears normal in carriers of a fragile X premutation.</a> <em style='font-style: italic'>Nat Genet</em>. <b>4</b>, 335-340 (1993).</li><li>Mullen, R. J., Buck, C. R., Smith, A. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NeuN,+a+neuronal+specific+nuclear+protein+in+vertebrates.">NeuN, a neuronal specific nuclear protein in vertebrates.</a> <em style='font-style: italic'>Development</em>. <b>116</b>, 201-211 (1992).</li><li>Sternberger, L. A., Sternberger, N. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Monoclonal+antibodies+distinguish+phosphorylated+and+nonphosphorylated+forms+of+neurofilaments+in+situ.">Monoclonal antibodies distinguish phosphorylated and nonphosphorylated forms of neurofilaments in situ.</a> <em style='font-style: italic'>Proc Natl Acad Sci U S A</em>. <b>80</b>, 6126-6130 (1983).</li></ol>

<p class="jove_content">在此过程中有两个关键的步骤，使大规模蛋白质的检测可能在整个猴脑。一个是嵌入和切片的协议，这仍然是国家安全局的专有知识。另一种是使用染色HistoTools提供的菜肴和篮子。后者可以方便，迅速地处理许多在某一特定时间（〜40）部分。它还提供了整个大脑的所有部分统一处理手段，使一个科学合理的组织学治疗。此外，嵌入式的标志性建筑的使用提供了额外的优势，能够以数字方式获取?...

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<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>anti-FMRP monoclonal antibody</td>
<td>&nbsp;</td>
<td>Chemicon</td>
<td>MAB2160</td>
<td>Requires antigen retrieval in fixed tissue.</td>
</tr>
<tr>
<td>anti-NeuN monoclonal antibody</td>
<td>&nbsp;</td>
<td>Chemicon</td>
<td>MAB377</td>
<td>N/A</td>
</tr>
<tr>
<td>anti-Neurofilament H monoclonal antibody</td>
<td>&nbsp;</td>
<td>Sternberger Monoclonals Inc.</td>
<td>SMI32</td>
<td>N/A</td>
</tr>
<tr>
<td>Antigen Unmasking Solution</td>
<td>&nbsp;</td>
<td>Vector Laboratories</td>
<td>H-3300</td>
<td>N/A</td>
</tr>
<tr>
<td>Normal horse serum</td>
<td>&nbsp;</td>
<td>Invitrogen</td>
<td>16050122</td>
<td>500 mL</td>
</tr>
<tr>
<td>Biotinylated Anti-Mouse IgG (H+L), made in horse</td>
<td>&nbsp;</td>
<td>Vector Laboratories</td>
<td>BA-2000</td>
<td>&nbsp;</td>
<tr>
<td>VECTASTAIN ABC Kit (Standard)</td>
<td>&nbsp;</td>
<td>Vector Laboratories</td>
<td>PK-4000</td>
<td>1.5 mg</td>
</tr>
<tr>
<td>Staining dish for floating sections</td>
<td>&nbsp;</td>
<td>HistoTools</td>
<td>10009</td>
<td>12 x 65 mm</td>
</tr>
<tr>
<td>Staining transport basket</td>
<td>&nbsp;</td>
<td>HistoTools</td>
<td>10012</td>
<td>large<br />
(fits 12 x 65 mm dish)</td>
</tr>
<tr>
<td>Triton X-100</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>P8514B</td>
<td>N/A</td>
</tr>
<tr>
<td>DAB</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>AC11209-0050</td>
<td>N/A</td>
</tr>		</tbody>
		</table>
		</div>
	

batch immunostaining for large-scale protein detection in the whole monkey brain

<p class="jove_content">免疫组织化学（IHC）是使用最广泛的实验室技术的一个靶蛋白的检测<em&gt;原位</em&gt;。了整个大脑的靶蛋白的表达模​​式的问题，比较容易回答的小脑筋，如啮齿动物的免疫组化使用时。然而，在大，令人费解的大脑，这些灵长类动物，如回答同样的问题提出了一些挑战。在这里，我们目前在成年猴脑的靶蛋白的免疫检测系统的方法。这种方法依赖于组织嵌入和神经科学协会（NSA）的切片方法以及自由浮动的部分批次染色专门开发的工具。结果部分，在一个特定的时间间隔，代表了整个大脑的染色均匀。造成的染色切片，可以经受各种各样的分析程序，以衡量蛋白质水平，表达某种蛋白的神经元的人口。</p>

Watch this Scientific Journal Video about Batch Immunostaining for Large-Scale Protein Detection in the Whole Monkey Brain at JoVE.com

Batch Immunostaining for Large-Scale Protein Detection in the Whole Monkey Brain

<p class="jove_content">大型整个灵长类动物大脑的靶蛋白的免疫检测是采用新颖的组织包埋，切片与创意仪器在某一特定时间使用多个自由浮动的部分批次的染色相结合的方法。</p>

在整个猴脑进行大规模蛋白质检测的批次免疫染色

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