Name: isolation of cd4+ t cells from mouse lymph nodes using miltenyi macs purification
Uploaded: 2007-11-01T00:34:55
Description: Watch this Scientific Journal Video about Isolation of CD4+ T cells from Mouse Lymph Nodes Using Miltenyi MACS Purification at JoVE.com

<p>我们要感谢编写的Mac缓冲丽贝卡Paquett。 </p>

<p>请阅读并遵守协议<a href="http://www.miltenyibiotec.com/">http://www.miltenyibiotec.com/</a>包括在美天旎套件或在网上。 </p>

<ol>
	<li>Miller, M. J., Hejazi, A. S., Wei, S. H., Cahalan, M. D., Parker, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=T+cell+repertoire+scanning+is+promoted+by+dynamic+dendritic+cell+behavior+and+random+T+cell+motility+in+the+lymph+node.">T cell repertoire scanning is promoted by dynamic dendritic cell behavior and random T cell motility in the lymph node.</a> <em style='font-style: italic'>Proc Natl Acad Sci U S A</em>. <b>101</b>, 998-1003 (2004).</li><li>Matheu, M. P., Deane, J. A., Parker, I., Fruman, D. A., Cahalan, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Class+IA+phosphoinositide+3-kinase+modulates+basal+lymphocyte+motility+in+the+lymph+node.">Class IA phosphoinositide 3-kinase modulates basal lymphocyte motility in the lymph node.</a> <em style='font-style: italic'>J Immunol</em>. <b>179</b>, 2261-2269 (2007).</li></ol>

<p>从主要来源细胞的分离是必要的步骤，在许​​多更复杂的协议。美天旎提供的工具包，隔离从几个生物体的细胞，包括人类在内的非人类灵长类动物，老鼠，我们在这里描述，老鼠。基于磁珠细胞分离，使无论是正面的选择（或细胞耗竭）以及阴性选择。在这里，我们展示了不变或NA已经CD4 +辅助性T细胞的阴性选择。使用这个标准的协议，我们通常净化细胞≥96％纯的CD4 + / CD3 +。虽然我们并没有表现出在这个?...

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		<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>MACS Buffer</td>
<td>Reagent</td>
<td>None</td>
<td>None </td>
<td>Prepare as described by Miltenyi. 

</td>
</tr>
<tr>
<td>CD4+ T cell Isolation Kit: Untouched Isolation </td>
<td>Reagent</td>
<td>Miltenyi</td>
<td>130-090-860</td>
<td>For use with the Miltenyi system. </td>
</tr>
<tr>
<td>Sterile Phosphate-Buffered Saline (PBS)</td>
<td>Reagent</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>For use in the preparation of MACS buffer.</td>
</tr>
<tr>
<td>Corning 35mm Not TC-Treated Culture Dish</td>
<td>Other</td>
<td>Corning Cell Culture Surfaces</td>
<td>430588</td>
<td>Used in the preparation of a single cell suspension.</td>
</tr>
<tr>
<td>70 micron Cell Strainer </td>
<td>Tool </td>
<td>BD Falcon</td>
<td>352350</td>
<td>Used in the preparation of a single cell suspension.</td>
</tr>
<tr>
<td>6 mL Syringe </td>
<td>Tool</td>
<td>Tyco Healthcare</td>
<td>8881516051</td>
<td>Used in the preparation of a single cell suspension.</td>
</tr>
<tr>
<td>10x Dulbecco's Phosphate Buffered Saline (DPBS)</td>
<td>Reagent</td>
<td>Invitrogen</td>
<td>14200-075</td>
<td>For use in RBC lysis.</td>
</tr>
<tr>
<td>Sterile Water </td>
<td>Reagent </td>
<td>Sigma</td>
<td>W4502</td>
<td>For use in RBC lysis.</td>
</tr>		</tbody>
		</table>
		</div>
			<div>
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isolation of cd4+ t cells from mouse lymph nodes using miltenyi macs purification

<p class="jove_content">从主要来源细胞的分离是必要的步骤，在许​​多更复杂的协议。美天旎提供的工具包，隔离从几个生物体的细胞，包括人类在内的非人类灵长类动物，老鼠，我们在这里描述，老鼠。基于磁珠细胞分离，使无论是正面的选择（或细胞耗竭）以及阴性选择。在这里，我们展示了不变或NA已经CD4 +辅助性T细胞的阴性选择。使用这个标准的协议，我们通常净化细胞≥96％纯的CD4 + / CD3 +。该协议是在与协议夹层和出版发行的朱庇特7，纯化的T细胞和其他类型的细胞过继转移成像目的，小鼠的外周淋巴结的双光子成像一起使用。虽然我们并没有表现出在这个协议的视频的流式细胞仪分析，它是强烈建议使用相应的抗体，通过流式细胞仪分离出的细胞检查的整体纯度。此外，我们证明T细胞隔离的非无菌的方法。如果无菌细胞是为您的特定最终用户的应用需要，一定要做好标准的无菌条件下，在组织培养罩中表现出的程序。感谢您看运气好，并用自己的实验！ </p>

Watch this Scientific Journal Video about Isolation of CD4+ T cells from Mouse Lymph Nodes Using Miltenyi MACS Purification at JoVE.com

Isolation of CD4+ T cells from Mouse Lymph Nodes Using Miltenyi MACS Purification

<p class="jove_content">使用美天旎互助委员会套件淋巴细胞分离是一种可靠的方法来净化整个淋巴组织匀浆的细胞。使用美天旎系统纯化的细胞通常是纯≥96％。在这里，我们展示了隔离CD4 + T细胞，由美天旎提供的许多工具包之一所采取的步骤。</p>

分离的CD4 +从小鼠淋巴节点使用美天旎MACS纯化的T细胞

Isolation of cells from the primary source is a necessary step in many more complex protocols. Miltenyi offers kits to isolate cells from several ...

Research

JoVE Journal

Biology

Dissection and 2-Photon Imaging of Peripheral Lymph Nodes in Mice

夹层和小鼠的外周淋巴结的双光子成像

Two-photon imaging has revealed an elegant choreography of motility and cellular interactions within the lymph node under basal conditions and at the ...

科研

生物学

CD4+ T-Lymphocyte Capture Using a Disposable Microfluidic Chip for HIV

CD4 + T淋巴细胞艾滋病毒的捕获使用一次性微流体芯片

Preparation of Single-Cell Suspensions from Mouse Spleen with the gentleMACS Dissociator

gentleMACS Dissociator从小鼠脾脏单细胞悬浮液的制备

Single-cell suspensions are a prerequisite for experiments in cell separation, cell analysis and cell culture.  To avoid tedious and often painful manual ...

Detection and Isolation of Viable Mouse IL-17-Secreting T Cells

可行的小鼠IL - 17分泌性T细胞的检测和隔离

The MACS Cytokine Secretion Assay technology allows detection of secreted cytokines on the single cell level and sensitive isolation of viable ...

Visualizing Antigen Specific CD4+ T Cells using MHC Class II Tetramers

可视化抗原特异性CD4 + T细胞的MHC II类四聚四

Major histocompatibility complex (MHC) class II tetramers allow the direct visualization of antigen specific CD4+ T cells by flow cytometry.  This method ...

Standardized Preparation of Single-Cell Suspensions from Mouse Lung Tissue using the gentleMACS Dissociator

标准化的制备小鼠肺组织单细胞悬浮液使用gentleMACS Dissociator

The preparation of single-cell suspensions from tissues is an important prerequisite for many experiments in cellular research. The process of ...

Generation of Single-Cell Suspensions from Mouse Neural Tissue

从小鼠神经组织产生单细胞悬浮液

Within the nervous system, hundreds of neuronal and glial cell types have been described. Each specific cell type in the brain or spinal cord has a ...

Purification of Mitochondria from Yeast Cells

从酵母细胞线粒体的纯化

Mitochondria are the main site of ATP production during aerobic metabolism in eukaryotic non-photosynthetic cells1. These complex organelles also play ...

Isolation of Valvular Endothelial Cells

瓣膜内皮细胞的分离

Heart valves are solely responsible for maintaining unidirectional blood flow through the cardiovascular system.  These thin, fibrous tissues are ...

Isolation of Retinal Stem Cells from the Mouse Eye

从鼠眼视网膜干细胞的分离

The adult mouse retinal stem cell (RSC) is a rare quiescent cell found within the ciliary epithelium (CE) of the mammalian eye1,2,3. The CE is made up of ...