Name: pouring and running a protein gel by reusing commercial cassettes
Uploaded: 2012-02-12T14:00:00
Description: Watch this Scientific Journal Video about Pouring and Running a Protein Gel by reusing Commercial Cassettes at JoVE.com

作者感谢用友科学为生命奖霍华德·休斯医学研究所的啊

1。准备磁带<ol><li>清洁的正面和背面板使用Invitrogen公司Nupage NOVEX微小胶体录像带，或其他商业微小胶体录像带。 </li><li>准备约1-3毫升的塑料环氧树脂，并根据制造商的指示，拌匀。 </li><li>适用于环氧树脂内缘的前面板上，两个板块接触时，磁带装配。 </li><li>通过背面板的底部缝滑入了一块过滤纸或纸板，装配时的录像带，以防止多余的环氧密封开幕。取出滤纸，只要这两个板块组装。 </li><li...

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	<li>Laemmli, U. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cleavage+of+structural+proteins+during+the+assembly+of+the+head+of+bacteriophage+T4.">Cleavage of structural proteins during the assembly of the head of bacteriophage T4.</a> Nature. 227, 680-685 (1970).</li><li>Raymond, S., Weintraub, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acrylamide+gel+as+a+supporting+medium+for+zone+electrophoresis.">Acrylamide gel as a supporting medium for zone electrophoresis.</a> Science. 130, 711-711 (1959).</li><li>Sch&#228;gger, H., von Jagow, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tricine-sodium+dodecyl+sulfate-polyacrylamide+gel+electrophoresis+for+the+separation+of+proteins+in+the+range+from+1+to+100+kDa.">Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa.</a> Anal. Biochem. 166, 368-379 (1987).</li><li>Shapiro, A. L., Vi&#241;uela, E., Maizel, J. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+weight+estimation+of+polypeptide+chains+by+electrophoresis+in+SDS-polyacrylamide+gels.">Molecular weight estimation of polypeptide chains by electrophoresis in SDS-polyacrylamide gels.</a> Biochem. Biophys. Res. Commun. 28, 815-820 (1967).</li></ol>

蛋白质凝胶电泳是大多数分子生物学实验室不可或缺的方法。市售minigels提供了方便的较高水平，但可能是昂贵的。在这里，我们展示如何重新使用从的​​市售minigels的流行品牌的微小胶体录音带。此方法保留了方便，但有一​​个现成的凝胶源的市售minigels成本的一小部分。在此过程中的一个关键步骤是运用正确的金额环氧树脂完全密封录音带之前加入unpolymerized丙烯酰胺，以防止泄漏。只使用...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> Invitrogen公司的Nupage的NOVEX微小胶体纸盒（1.0毫米） </td><td> Invitrogen公司</td><td> NC2010 </td><td>可在2，5，10，12，15和2D井</td></tr><tr><td>塑料环氧树脂</td><td>乐泰</td><td> 108771 </td><td></td></tr><tr><td>电工胶带</td><td>苏格兰威士忌</td><td></td><td></td></tr><tr><td> 125毫升侧臂瓶</td><td>耐热</td><td> 5360 </td><td></td></tr><tr><td> 4X Tris碱分离胶缓冲</td><td></td><td></td><td> 1.5 Tris碱的pH值8.7 </td></tr><tr><td>丙烯酰胺</td><td> Fisher Scientific则</td><td> 01065-500 </td><td></td></tr><tr><td> BIS-丙烯酰胺</td><td> MP Biomedicals， </td><td> 800173 </td><td></td></tr><tr><td> 30.8％的丙烯酰胺溶液</td><td></td><td></td><td> 30:0.8丙烯酰胺：双丙烯酰胺表意文字-H 2 O </td></tr><tr><td> 10％的APS </td><td> Fisher Scientific则</td><td> BP179-25 </td><td> 0.1克1毫升表意文字- H 2 O的过硫酸铵，立即使用前准备</td></tr><tr><td> TEMED </td><td> Arcos的有机物</td><td> 138450500 </td><td></td></tr><tr><td> 1 - 丁醇饱和1X分离胶缓冲</td><td></td><td></td><td> 5:2 1 - 丁醇1X分离胶缓冲</td></tr><tr><td> 4X Tris碱浓缩胶缓冲</td><td></td><td></td><td> 1.5 Tris碱pH值6.8 </td></tr><tr><td>凝胶梳</td><td> Invitrogen公司</td><td> NC3015 </td><td> 15好，1.0毫米厚度</td></tr><tr><td>脉冲封口机</td><td>趋势TEW电暖气设备</td><td></td><td></td></tr><tr><td>热封小包</td><td> kapak / Scotchpak </td><td></td><td> 1品脱大小（6.5“×8”） </td></tr><tr><td>凝胶存储缓冲区</td><td></td><td></td><td> 1X叠加0.1％叠氮化钠凝胶缓冲</td></tr><tr><td> NOVEX SDS样品缓冲液Tris-甘氨酸（2X） </td><td> Invitrogen公司</td><td> LC2676 </td><td></td></tr><tr><td>数码地面电视</td><td> FisherBiotech </td><td> BP172-5 </td><td></td></tr><tr><td> 10X的三Gylcine跑缓冲</td><td> Invitrogen公司</td><td> LC2675 </td><td> 10X股票1L：29.0Ğ三基地，144.0 g甘氨酸，10.0ĞSDS </td></tr><tr><td> Xcell的SureLock迷你细胞</td><td> Invitrogen公司</td><td> EI0001 </td><td></td></tr><tr><td>胶刀</td><td> Invitrogen公司</td><td> EI9010 </td><td></td></tr><tr><td> SimplyBlue安全染色</td><td> Invitrogen公司</td><td> LC6065 </tD&gt; <td></td></tr></tbody></table>

pouring and running a protein gel by reusing commercial cassettes

使用十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS - PAGE）分析蛋白质的评价是一种常见的技术，生物化学和分子生物学的研究人员1-4。执行每日蛋白质分析实验室，市售的聚丙烯酰胺凝胶（$ 10/gel）的成本可能是相当一段时间。为了降低成本，一些研究人员自己编写的聚丙烯酰胺凝胶。这些凝胶浇注的传统方法通常利用专门的设备和玻璃的凝胶板，可以是昂贵的，排除许多凝胶浇注和存储以供将来使用。此外，在清洗或浇注凝胶玻璃板处理可能会导致意外破损，创造一个安全隐患，这可能妨碍其使用在本科实验班。我们的协议演示如何同时倒入多个蛋白凝胶回收Invitrogen公司Nupage NOVEX微小胶体录音带，和廉价的马terials家装店购买。这种经济和精简的方法，包括了几个星期的方式储存在4°C凝胶。通过重新使用塑料胶盒从市售凝胶，实验室运行频繁的蛋白凝胶可显着节省成本，并帮助环境。此外，塑料胶录音带是极耐破损，这使得他们的本科生做实验教室的理想。

Watch this Scientific Journal Video about Pouring and Running a Protein Gel by reusing Commercial Cassettes at JoVE.com

Pouring and Running a Protein Gel by reusing Commercial Cassettes

我们的协议演示如何倒在时间的多种蛋白质凝胶回收Invitrogen公司Nupage NOVEX微小胶体录音带，并在家装商店购买廉价的材料。这种经济和精简的方法，包括了几个星期的方式储存在4°C凝胶。通过重新使用塑料胶盒从市售凝胶，实验室运行频繁的蛋白凝胶可显着节省成本，并帮助环境。

浇注和运行的一种蛋白质凝胶重用商​​业磁带

The evaluation of proteins using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis is a common technique used by biochemistry ...

Research

JoVE Journal

Biology

Denaturing Gradient Gel Electrophoresis (DGGE)

Denaturing Gradient Gel Electrophoresis DGGE

变性梯度凝胶电泳（DGGE）

科研

生物学

Screening for Amyloid Aggregation by Semi-Denaturing Detergent-Agarose Gel Electrophoresis

半变性洗涤剂琼脂糖凝胶电泳筛选淀粉样蛋白聚合

Amyloid aggregation is associated with numerous protein misfolding pathologies and underlies the infectious properties of prions, which are ...

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

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 ...

Using the GELFREE 8100 Fractionation System for Molecular Weight-Based Fractionation with Liquid Phase Recovery

液相恢复GELFREE 8100分馏系统使用基于分子量分馏

The GELFREE 8100 Fractionation System is a novel protein fractionation system designed to maximize protein recovery during molecular weight based ...

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

Isolation of Labile Multi-protein Complexes by in vivo Controlled Cellular Cross-Linking and Immuno-magnetic Affinity Chromatography

由不稳定的多蛋白质复合物的分离在体内控制细胞的交联和免疫磁性亲和层析

The dynamic nature of cellular machineries is frequently built on transient and/or weak protein associations. These low affinity interactions preclude ...

Orthogonal Protein Purification Facilitated by a Small Bispecific Affinity Tag

由一个小型的双特异性亲和标签的推动正交蛋白质纯化

Due to the high costs associated with purification of recombinant proteins the protocols need to be rationalized. For high-throughput efforts there is a ...

Improving the Success Rate of Protein Crystallization by Random Microseed Matrix Screening

提高蛋白质结晶的成功率随机Microseed矩阵筛选

Random microseed matrix screening  (rMMS) is a protein crystallization technique in which seed crystals are added  to random screens. By increasing the ...

Protein-protein Interactions Visualized by Bimolecular Fluorescence Complementation in Tobacco Protoplasts and Leaves

蛋白质 - 蛋白质相互作用的双分子荧光互补的烟草原生质体和叶片可视化

Many proteins interact transiently with other proteins or are integrated into multi-protein complexes to perform their biological function. Bimolecular ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

在造血干细胞和祖细胞的成年小鼠颅骨骨髓体内四维跟踪

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

Green Fluorescent Protein-based Expression Screening of Membrane Proteins in Escherichia coli

膜蛋白的绿色荧光蛋白为基础的表达筛选<em&gt;大肠杆菌</em

The production of recombinant membrane proteins for structural and functional studies remains technically challenging due to low levels of expression and ...

浇注和运行的一种蛋白质凝胶重用商​​业磁带

浇注和运行的一种蛋白质凝胶重用商业磁带