Name: staining of proteins in gels with coomassie g-250 without organic solvent and acetic acid
Uploaded: 2009-08-14T00:00:00
Description: Watch this Scientific Journal Video about Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid at JoVE.com

<p class="jove_content"> ，我们愿意承认的伊内斯拉克的技术援助。 </p>

<p class="jove_title">第1部分：CBB染色溶液的制备</p><ol><li> 60-80毫克CBB G - 250是在1升bidistilled水溶解，搅拌2-4小时。最后，3毫升浓盐酸加入搅拌一分钟，以深蓝色的解决方案，并存储在黑暗供日后使用。该解决方案可存储几个星期到几个月的时间，而不会失去其染色效率。 </li><li>浓盐酸应处理与常规护理UND通风​​柜下使用。最终的解决方案将在pH为2左右，所以手套，应使用和任何应避免与皮肤接触。 </li></ol><p class="j...

<ol>
	<li>Wondrak, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Process+for+fast+visualization+of+protein.">Process for fast visualization of protein.</a> <em style='font-style: italic'>US patent</em>. , (2001).</li><li>Wondrak, E. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Solution+for+fast+visualization+of+protein.">Solution for fast visualization of protein.</a> <em style='font-style: italic'>US patent</em>. , (2001).</li></ol>

<ul><li>洗涤步骤是为高效的蛋白质染色是至关重要的。 2分以下或水（&lt;50毫升）的体积缩小减少洗涤时间，可以在淡蓝色的蛋白条带，最有可能由于凝胶中残留的SDS的结果。 </li><li>如果蛋白质质谱分析，在微波炉中加热的步骤应跳过，扩展到约10分钟，每一步和染色时间延长，直到带的强度凝胶洗涤时间足够强大。加热质谱的蛋白质的交联凝胶的凝胶基质，从而检测的蛋白质可能会受到阻碍。 </li><li>相反的CBB G - ...

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<td>Coomassie Brilliant Blue G-250</td>
<td>&nbsp;</td>
<td>AppliChem</td>
<td>A3480</td>
<td>any other CBB G-250 could be used as well</td>
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<td>Concentrated HCl</td>
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staining of proteins in gels with coomassie g-250 without organic solvent and acetic acid

<p class="jove_content">在古典蛋白染色用考马斯亮蓝（CBB）与有毒和易燃的有机溶剂（甲醇，乙醇或异丙醇）和乙酸含量高的解决方案的协议是用于固定，染色和脱色凝胶中的蛋白质经SDS - PAGE分析。为了加快程序，在很短的时间微波炉加热的染色溶液是常用的。这通常会导致有毒或有害的甲醇，乙醇或异丙醇和醋酸在实验室应避免由于安全方面的考虑的强烈气味的挥发。在最初两个EM Wondrak（US2001046709（A1），US6319720（B1））专利申请公布一个协议，染色液的另一种成分是其中不含有机溶剂或酸是用来描述。溶解在bidistilled水的CBB（CBB G - 250每公升60 - 80毫克）和35毫米盐酸是作为唯一的染色溶液中的其他化合物的补充。 CBB staning凝胶完成后bidistilled水凝胶的SDS - PAGE和彻底清洗。通过加热凝胶在洗涤和染色步骤，完成这个过程可以更快，任何有毒或有害compunds蒸发。染色的蛋白质发生已经在1分钟内加热后的凝胶染色溶液和全面发展与略带蓝色的背景，完全是destained长时间洗染色bidistilled水凝胶后15-30分钟，而不会影响染色的蛋白质，带。</p>

Watch this Scientific Journal Video about Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid at JoVE.com

Staining of Proteins in Gels with Coomassie G-250 without Organic Solvent and Acetic Acid

<p class="jove_content">蛋白质与考马斯亮蓝（CBB）的G - 250在聚丙烯酰胺凝胶染色的一个短的协议是在古典与CBB染色程序，而无需使用有机溶剂或醋酸。</p>

无有机溶剂和醋酸，蛋白质与考马斯亮蓝G - 250凝胶染色

In classical protein staining protocols using Coomassie Brilliant Blue (CBB), solutions with high contents of toxic and flammable organic solvents ...

Research

JoVE Journal

Biology

Staining Proteins in Gels

在凝胶染色蛋白质

Following separation by electrophoretic methods, proteins in a gel can be detected by several staining methods. This unit describes protocols for ...

科研

生物学

Fast and Sensitive Colloidal Coomassie G-250 Staining for Proteins in Polyacrylamide Gels

快速，灵敏的胶体考马斯亮G - 250染色蛋白聚丙烯酰胺凝胶

Coomassie Brilliant Blue (CBB) is a dye commonly used for the visualization of proteins separated by SDS-PAGE, offering a simple staining procedure and ...

Profiling of Methyltransferases and Other <em>S</em>-adenosyl-<sub>L</sub>-homocysteine-binding Proteins by Capture Compound Mass Spectrometry (CCMS)

Profiling of Methyltransferases and Other S-adenosyl-L-homocysteine-binding Proteins by Capture Compound Mass Spectrometry CCMS

分析甲基化和其他<em> S</em>腺苷<sub> L</sub捕捉复合质谱（CCMS）>同型半胱氨酸结合蛋白

There is a variety of approaches to reduce the complexity of the proteome on the basis of functional small molecule-protein interactions such as affinity ...

High-Throughput Measurement and Classification of Organic P in Environmental Samples

高通量测量和环境样品中的有机磷分类

Many types of organic phosphorus (P) molecules exist in environmental samples1.  Traditional P measurements do not detect these organic P compounds since ...

Synthesis of Nine-atom Deltahedral Zintl Ions of Germanium and their Functionalization with Organic Groups

九的原子Deltahedral Zintl离子锗和他们的官能合成有机基团

Although the first studies of Zintl ions date between the late 1890's and early 1930's they were not structurally characterized until many years later.1,2 ...

Genotyping of Plant and Animal Samples without Prior DNA Purification

基因分型的植物和动物样品，恕不另行DNA的纯化

The Direct PCR approach facilitates PCR amplification directly from small amounts of unpurified samples, and is demonstrated here for several plant and ...

Analyzing and Building Nucleic Acid Structures with 3DNA

核酸结构分析及建设3DNA

The 3DNA software package is a popular and versatile bioinformatics tool with capabilities to analyze, construct, and visualize three-dimensional nucleic ...

A Chemical Screening Procedure for Glucocorticoid Signaling with a Zebrafish Larva Luciferase Reporter System

对糖皮质激素信号A化学筛选程序与斑马鱼幼虫的荧光素酶报告系统

Glucocorticoid stress hormones and their artificial derivatives are widely used drugs to treat inflammation, but long-term treatment with glucocorticoids ...

Sequence-specific Labeling of Nucleic Acids and Proteins with Methyltransferases and Cofactor Analogues

核酸，蛋白质与甲基转移和辅酶类似物序列特异性标签

S-Adenosyl-l-methionine (AdoMet or SAM)-dependent methyltransferases (MTase) catalyze the transfer of the activated methyl group from AdoMet to specific ...

Visualization of miniSOG Tagged DNA Repair Proteins in Combination with Electron Spectroscopic Imaging (ESI)

Visualization of miniSOG Tagged DNA Repair Proteins in Combination with Electron Spectroscopic Imaging ESI

在联合电子能谱成像miniSOG标记的DNA修复蛋白可视化（ESI）

The limits to optical resolution and the challenge of identifying specific protein populations in transmission electron microscopy have been obstacles in ...