Name: measuring the 50% haemolytic complement (ch<sub>50</sub>) activity of serum
Uploaded: 2010-03-29T18:05:00
Description: Watch this Scientific Journal Video about Measuring the 50% Haemolytic Complement CH50 Activity of Serum at JoVE.com

<p class="jove_content">笔者想感谢劳拉马修斯小姐表演的技巧和摄影郑明牧羊犬。这个项目是由南澳大学，实验室医学程序。 </p>

<p class="jove_title">制备的5倍Veronal缓冲液（VBS） </p><ol><li>要准备Veronal缓冲液（VBS），3个单独的解决方案都需要做好准备。 </li><li>准备通过溶解在蒸馏水350毫升NaCl和0.94gm钠巴比妥21.25gm溶液1。氯化钠和巴比妥钠终浓度分别为1.02M和13MM。 </li><li>准备通过溶解在热蒸馏水125毫升巴比妥1.44gm溶液2。巴比妥最终浓度62.5mM。 </li><li>准备通过溶解在蒸馏水100ml <sub>氯化钙</sub>氯化镁<sub>2</sub>和4.41gm 20.33gm溶液3。 <sub>2</sub>氯化镁...

<ol>
	<li>Goldsby, R. A., Kindt, T. J., Osborne, B. A., Kuby, J. <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'>Immunology</em>. , (2003).</li></ol>

<p class="jove_content"> CH <sub>50</sub>含量是受到很多干扰。有些SRBC的是比其他人更脆弱，导致自发溶血补体活性无关。兔抗体的亲和力很多很多，从生产厂家之一到另一个不同，这影响的SRBC结合的抗体量。此外，敏与在细胞与不同数量的抗体涂层的SRBC的抗体结果SRBC的过程。标本采集和存储是一个重要的潜在的错误来源。补体成分如补体C1q，C3，C4和C5是非常不稳定的，因此适当的样品处理是至关重要的。长时间暴露在?...

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<td>Sheep red blood cells</td>
<td>&nbsp;</td>
<td>CSL</td>
<td>2490201</td>
<td>Use at 1% final</td>
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<tr>
<td>Serum</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Human serum</td>
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<td>Rabbit Anti-Sheep Haemolytic serum (RBCs), Unconjugated</td>
<td>&nbsp;</td>
<td>AdB Serotec</td>
<td>C12HSB</td>
<td>&nbsp;</td>
<tr>
<td>96 well flat bottom plate</td>
<td>&nbsp;</td>
<td>Sarstedt</td>
<td>83.1839</td>
<td>&nbsp;</td>
<tr>
<td>Veronal buffered saline</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Use at 1x final</td>
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<td>Waterbath</td>
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<td>Plate reader</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
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<td>37&deg;C room/incubator</td>
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measuring the 50% haemolytic complement (ch<sub>50</sub>) activity of serum

<p class="jove_content">补体系统是一组蛋白质，当激活，导致靶细胞裂解，促进吞噬通过opsonisation。个人补体成分是可以量化的，但这不提供任何信息的途径的活动。这架CH<sub&gt; 50</sub&gt;是激活经典补体途径（图1）筛选实验，它是敏感的任何组件的通路减少，缺乏和/或闲置。这架CH<sub&gt; 50</sub&gt;测试的经典途径裂解绵羊红细胞（SRBC），兔抗绵羊红细胞抗体（溶血）预涂血清补体成分的功能性。当抗体包被的SRBC培养与血清测试，激活补体经典途径和溶血结果。如果缺少的一个补充部分，在CH<sub&gt; 50</sub&gt;等级将为零;如果一个或多个组件的经典途径是减少，在CH<sub&gt; 50</sub&gt;会有所下降。一个固定体积的最佳致敏SRBC被添加到每个血清稀释。孵育后，将混合物离心和量化的溶血程度是通过测量血红蛋白释放到上清液于540nm处的吸光度。补体活性的金额是由研究能力的各种测试血清稀释溶解抗体包被的SRBC。此视频概述参与分析经典补体途径的补充活动水平的实验步骤。</p>

Watch this Scientific Journal Video about Measuring the 50% Haemolytic Complement CH50 Activity of Serum at JoVE.com

Measuring the 50% Haemolytic Complement CH50 Activity of Serum

<p class="jove_content">经典途径激活抗体和靶细胞裂解高潮。这架CH<sub&gt; 50</sub&gt;检测提供了一个补体活性的血清样本的措施。本视频演示的CH在确定所涉及的步骤<sub&gt; 50</sub&gt;血清样本，计算和解释结果。</p>

测量50％的溶血性补充（CH<sub> 50</sub>）活性的血清

The complement system is a group of proteins that when activated lead to target cell lysis and facilitates phagocytosis through opsonisation. Individual ...

Research

JoVE Journal

Immunology and Infection

Measuring the 50% Haemolytic Complement (CH<sub>50</sub>) Activity of Serum

Depletion of Specific Cell Populations by Complement Depletion

补消耗的特定细胞群的枯竭

The purification of immune cell populations is often required in order to study their unique functions.  In particular, molecular approaches such as ...

科研

免疫与感染

Determining the Reactivity and Titre of Serum using a Haemagglutination Assay

确定使用血凝检测血清中的反应和滴度

Haemagglutination is a specific form of agglutination and is used when antibodies bind to red blood cells, which act as a particulate antigen. Red blood ...

Measuring Calpain Activity in Fixed and Living Cells by Flow Cytometry

在固定和活细胞，流式细胞仪测量钙蛋白酶活性

Calpains are ubiquitous intracellular, calcium-sensitive, neutral cysteine proteases 1. Calpains play crucial roles in many physiological processes, ...

Measuring Bacterial Load and Immune Responses in Mice Infected with <em>Listeria monocytogenes</em>

Measuring Bacterial Load and Immune Responses in Mice Infected with Listeria monocytogenes

测量细菌感染小鼠和免疫反应<em>李斯特菌</em

Listeria monocytogenes (Listeria) is a Gram-positive facultative intracellular pathogen1. Mouse studies typically employ intravenous injection of ...

Determining the Phagocytic Activity of Clinical Antibody Samples

确定临床抗体样品的吞噬活性

Antibody-driven phagocytosis is induced via the engagement of Fc receptors on professional phagocytes, and can contribute to both clearance as well as ...

Saliva, Salivary Gland, and Hemolymph Collection from <em>Ixodes scapularis</em> Ticks

Saliva, Salivary Gland, and Hemolymph Collection from Ixodes scapularis Ticks

唾液，唾液腺，淋巴收集肩突硬蜱蜱

Ticks are found worldwide and afflict humans with many tick-borne illnesses. Ticks are vectors for pathogens that cause Lyme disease and tick-borne ...

Methods for Quantitative Detection of Antibody-induced Complement Activation on Red Blood Cells

定量检测抗体诱导补体激活的方法对红血细胞

Antibodies against red blood cells (RBCs) can lead to complement activation resulting in an accelerated clearance via complement receptors in the liver ...

Measuring Local Anaphylaxis in Mice

小鼠量度本地过敏反应

Allergic responses are the result of the activation of mast cells and basophils, and the subsequent release of vasoactive and proinflammatory mediators. ...

Measuring Granulocyte and Monocyte Phagocytosis and Oxidative Burst Activity in Human Blood

人体血液中测粒及单核细胞吞噬功能和呼吸爆发活动

The granulocyte and monocyte phagocytosis and oxidative burst (OB) activity assay can be used to study the innate immune system. This manuscript provides ...

Assays for Studying the Role of Vitronectin in Bacterial Adhesion and Serum Resistance

Vitronectin 在细菌黏附和血清耐药性中作用的实验研究

Bacteria utilize complement regulators as a means of evading the host immune response. Here, we describe protocols for evaluating the role vitronectin ...

测量50％的溶血性补充（CH 50）活性的血清

测量50％的溶血性补充（CH₅₀）活性的血清