Name: preparation of drosophila polytene chromosome squashes for antibody labeling
Uploaded: 2010-02-09T17:00:00
Description: Watch this Scientific Journal Video about Preparation of Drosophila Polytene Chromosome Squashes for Antibody Labeling at JoVE.com

我们感谢五Lephart女士维护飞生补助金（GM62916）和国家科学基金会的资助（MCB0817107），这项工作是由国家机构支持的股票。

聚乙烯染色体壁球准备以下协议是改编自约翰森等人所描述的程序。 （2009年）。 1。文化的第三龄果蝇幼虫为了获得高品质壁球制剂的最佳聚乙烯染色体，拥挤的培养条件是必不可少的（即大约20个鸡蛋在一个标准的4个“飞瓶和变化到一个新的瓶子铺设雌性果蝇每天），选择最胖的个人从第一次攀登，而他们仍然徘徊，但只是到化蛹前幼虫3?...

<ol>
	<li>Cai, W., Bao, X., Deng, H., Jin, Y., Girton, J., Johansen, J., Johansen, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=RNA+polymerase+II-mediated+transcription+at+active+loci+does+not+require+histone+H3S10+phosphorylation+in+Drosophila.">RNA polymerase II-mediated transcription at active loci does not require histone H3S10 phosphorylation in Drosophila.</a> Development. 135, 2917-2925 (2008).</li><li>DiMario, P., Rosby, R., Cui, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct+visualization+of+GFP-fusion+proteins+on+polytene+chromosomes.">Direct visualization of GFP-fusion proteins on polytene chromosomes.</a> Dros. Inf. Serv. 89, 115-118 (2006).</li><li>Johansen, K. M., Cai, W., Deng, H., Bao, X., Zhang, W., Girton, J., Johansen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Methods+for+studying+transcription+and+epigenetic+chromatin+modification+in+Drosophila+polytene+chromosome+squash+preparations+using+antibodies.">Methods for studying transcription and epigenetic chromatin modification in Drosophila polytene chromosome squash preparations using antibodies.</a> Methods. 48, 387-397 (2009).</li></ol>

包含在传统壁球固定协议的醋酸和乳酸，有利于带间的决议和染色体臂蔓延，但不幸的是，一些抗原决定簇不生存之本治疗。这样一个抗原表位的一个例子是H3S10ph（ 蔡等 ，2008） 。由于酸的治疗也有它的缺点是淬火GFP标记的蛋白质，DiMario等固有荧光。 （2006年）最近开发的基于甲醛的“无酸壁球技术”，允许无酸敏感的抗原决定簇的抗体检测GFP抗体标记以及GFP融合蛋白聚乙烯染色?...

preparation of drosophila polytene chromosome squashes for antibody labeling

果蝇一直喜爱的模型系统研究由于聚乙烯染色体三龄幼虫的巨大唾液腺提供细胞学优势的染色质结构和基因调控之间的关系。在这个组织中的染色体在细胞分工上升到约1000份的情况下进行多轮的复制。的DNA仍然大大扩大提供一个独特的机会，到相关的特定蛋白质的本地化染色质形态的染色体复制后每个周期相一致。因此，出现了一个高水平的界定目前在不同的基因的表观遗传修饰和转录过程的不同阶段的兴趣。为此类研究的重要工具，是与抗体的酶标记的聚乙烯染色体，转录因子，或感兴趣的组蛋白修饰。这个视频协议说明，在约翰森实验室用于制备抗体标记果蝇多线染色体的壁球技术。

Watch this Scientific Journal Video about Preparation of Drosophila Polytene Chromosome Squashes for Antibody Labeling at JoVE.com

Preparation of Drosophila Polytene Chromosome Squashes for Antibody Labeling

该视频协议说明在约翰森实验室中使用的壁球技术准备<em&gt;果蝇</em&gt;聚乙烯抗体标记染色体。

果蝇聚乙烯南瓜抗体标记染色体的制备

Drosophila has long been a favorite model system for studying the relationship between chromatin structure and gene regulation due to the cytological ...

Research

JoVE Journal

Biology

Preparation of Neuronal Cultures from Midgastrula Stage Drosophila Embryos

从Midgastrula舞台果蝇胚胎神经文化的制备

This video illustrates the procedure for making primary neuronal cultures from midgastrula stage Drosophila embryos. The methods for collecting embryos ...

科研

生物学

The Preparation of Drosophila Embryos for Live-Imaging Using the Hanging Drop Protocol

The Preparation of Drosophila Embryos for Live-Imaging Using the Hanging Drop Protocol

编制果蝇实时成像的胚胎使用悬滴协议

Green fluorescent protein (GFP)-based timelapse live-imaging is a powerful technique for studying the genetic regulation of dynamic processes such as ...

Fluorescent Labeling of Drosophila Heart Structures

Fluorescent Labeling of Drosophila Heart Structures

荧光标记的果蝇心结构

The Drosophila melanogaster dorsal vessel, or heart, is a tubular structure comprised of a single layer of contractile cardiomyocytes, pericardial cells ...

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

电子显微镜对胚胎的制备果蝇胚胎心管

The morphogenesis of the Drosophila  embryonic heart tube has emerged as a valuable model system for studying cell migration, cell-cell adhesion and cell ...

Live Dissection of Drosophila Embryos: Streamlined Methods for Screening Mutant Collections by Antibody Staining

Live Dissection of Drosophila Embryos: Streamlined Methods for Screening Mutant Collections by Antibody Staining

现场剖析果蝇胚胎抗体染色筛选突变体集合的简化方法

Drosophila embryos between stages 14 and 17 of embryonic development can be readily dissected to generate "fillet" preparations.  In these preparations, ...

Preparation of Developing and Adult Drosophila Brains and Retinae for Live Imaging

Preparation of Developing and Adult Drosophila Brains and Retinae for Live Imaging

发展和成人的制备果蝇大脑和视网膜实时成像

The Drosophila brain and visual system are widely utilized model systems to study neuronal development, function and degeneration. Here we show three ...

Preparation of Drosophila S2 cells for Light Microscopy

Preparation of Drosophila S2 cells for Light Microscopy

制备果蝇 S2细胞，光学显微镜

The ideal experimental system would be cheap and easy to maintain, amenable to a variety of techniques, and would be supported by an extensive literature ...

Preparation of Adult Drosophila Eyes for Thin Sectioning and Microscopic Analysis

Preparation of Adult Drosophila Eyes for Thin Sectioning and Microscopic Analysis

成人的制备果蝇</em薄切片和显微镜分析>的眼睛

Drosophila has long been used as model system to study development, mainly due to the ease with which it is genetically tractable. Over the years, a ...

Studying Mitotic Checkpoint by Illustrating Dynamic Kinetochore Protein Behavior and Chromosome Motion in Living Drosophila Syncytial Embryos

Studying Mitotic Checkpoint by Illustrating Dynamic Kinetochore Protein Behavior and Chromosome Motion in Living Drosophila Syncytial Embryos

龙虎斗动粒蛋白的动态行为与染色体运动，在生活学习有丝分裂关卡<em&gt;果蝇</em&gt;合胞胚胎

The spindle assembly checkpoint (SAC) mechanism is an active signal, which monitors the interaction between chromosome kinetochores and spindle ...

Chromosome Preparation From Cultured Cells

染色体制备培养细胞

Chromosome (cytogenetic) analysis is widely used for the detection of chromosome instability. When followed by G-banding and molecular techniques such as ...