Name: Capturando quebras de local frágeis comuns por γH2A nativo. X ChIP (Vídeo) | JoVE Journal
Uploaded: 2025-01-24T14:20:00.000+00:00
Duration: 9 min 46 s
Description: Aprenda a detectar quebras em sítios frágeis comuns usando ChIP nativo de γH2A.X. Este método rápido e eficiente reduz o tempo de trabalho em pesquisas sobre instabilidade genômica em células cancerígenas.

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<li>Miyamoto, R., Yokoyama, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Protocol+for+fractionation-assisted+native+ChIP+%28fanChIP%29+to+capture+protein-protein%2FDNA+interactions+on+chromatin%5BTitle%5D)+AND+%22STAR+Protoc%22%5BJournal%5D)">Protocol for fractionation-assisted native ChIP (fanChIP) to capture protein-protein/DNA interactions on chromatin.</a> STAR Protoc. 2 (2), 100404(2021).</li>
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</ol>

<table><tbody><tr><td>0,2 &micro; membrana de nitrocelulose m</td><td>Amersham</td><td>10600011</td><td></td></tr><tr><td>Actina B</td><td>ProteinTech</td><td>20536-1-AP</td><td></td></tr><tr><td>Afidicolina</td><td>MedChemExpress</td><td>HY-N6733</td><td></td></tr><tr><td>Grânulos magnéticos de proteína A / G de grau ChIP</td><td>ThermoFisher</td><td>26162</td><td></td></tr><tr><td>Substrato Clarity Western ECL</td><td>Bio-Rad</td><td>#1705061</td><td></td></tr><tr><td>Glicogênio, grau de biologia molecular</td><td>ThermoFisher</td><td>Gato. Não.&nbsp; R0561</td><td></td></tr><tr><td>Anticorpo secundário conjugado com HRP&nbsp;</td><td>ProteinTech</td><td>SA00001-2</td><td></td></tr><tr><td>hidroxiureia&nbsp;</td><td>MedChemExpress</td><td>HY-B0313</td><td></td></tr><tr><td>Nuclease Microcócica&nbsp;</td><td>NEB</td><td>M0247S</td><td></td></tr><tr><td>IgG normal&nbsp;</td><td>Santa Cruz</td><td>SC-2025</td><td></td></tr><tr><td>Taq Universal SYBR Green Supermix&nbsp;</td><td>BioRad</td><td>1725120</td><td></td></tr><tr><td>&gama; H2A. anticorpo X&nbsp; (para ChIP)</td><td>Sigma-Aldrich</td><td>05-636</td><td></td></tr><tr><td>&gama; H2A. X (para WB)</td><td>Sinalização Celular</td><td>#25955</td><td></td></tr></tbody></table>

capturando quebras de local frágeis comuns por γh2a nativo. x chip

Capturing Common Fragile Site Breaks by Native &#947;H2A.X ChIP

O tamanho dos fragmentos de cromatina é crucial para o sucesso do Native ChIP, pois afeta diretamente a acessibilidade das regiões de DNA para ligação de anticorpos. Para determinar a concentração ideal de MNase para fragmentação da cromatina, preparamos uma série de tubos de microcentrífuga contendo concentrações variadas de MNase (ou seja, 0,0625 U, 0,125 U, 0,25 U, 0,5 U, 1 U, 2 U, 4 U, 8 U por reação) e 40 μL de núcleos isolados. Cada reação foi incubada a 37 ° C por 5 min para atingir uma variedade de tamanhos de fr...

Capturando quebras de local frágeis comuns por γH2A nativo. X ChIP

Replication stress induced by exposure to extrinsic agents can lead to DNA breaks at common fragile sites, which are regions in the genome known to be ...

capturing-common-fragile-site-breaks-by-native-h2ax-chip

Research

JoVE Journal

Biochemistry

Bioquímica

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Contínua observação microscópica de alta resolução de replicativa envelhecimento em leveduras de brotamento

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A ruptura dielétrica dependente do tempo (TDDB) em pilhas de interconexão no chip é um dos mecanismos de falha mais críticos para dispositivos microeletrônicos. Este trabalho demonstra o procedimento de um experimento TDDB in situ no microscópio eletrônico de transmissão, que abre a possibilidade de estudar o mecanismo de falha em produtos microeletrônicos.

Em ruptura dielétrica Situ dependentes do tempo no Microscópio Eletrônico de Transmissão: A possibilidade de compreender o mecanismo de falha em dispositivos microeletrônicos

Relatamos um protocolo para combinar a metrologia atômica do Microscópio de Tunelamento de Varredura para padronização de superfície com Deposição Atômica Seletiva de Camada e Gravação de Íons Reativos. Usando um processo robusto envolvendo inúmeras exposições atmosféricas e transporte, nanoestruturas 3D com metrologia atômica são fabricadas.

Atomicamente rastreável Fabrication Nanostructure

Aqui apresentamos os protocolos experimentais para a observação em tempo real de um processo Self-assembly usando microscopia eletrônica de transmissão de células líquido.

Microscopia eletrônica de transmissão de líquido-celular para rastreamento auto-montagem de nanopartículas

Aqui, apresentamos um protocolo que descreve a fabricação do stretchable, dual channel, dispositivos de cultura de célula órgão microplaqueta microfluidic para recapitulando a funcionalidade de nível de órgão em vitro.

Fabricação escalável de Stretchable, Dual Channel, órgão Microfluidic Chips

Este artigo descreve um protocolo juntamente com um estudo comparativo de duas técnicas de fabricação microfluida, a fotolitografia/gravura úmida/ligação térmica e Gravura Seletiva induzida por laser (SLE), que são adequadas para condições de alta pressão. Essas técnicas constituem plataformas de habilitação para observação direta do fluxo de fluidos em mídia permeável substituta e sistemas fraturados em condições de reservatório.

Técnicas de fabricação microfluidica para testes de alta pressão do transporte de espuma de CO2 supercrítica de microescala em reservatórios não convencionais fraturados

Aqui, apresentamos um protocolo para a realização in situ TEM experimentos de reação de gás de células fechadas enquanto detalhamos vários métodos de preparação de amostras comumente usados.

Realizando reações de gás de célula fechada situ no microscópio eletrônico de transmissão

Um protocolo para a cultura e manipulação do tecido embrionário do rato em um chip microfluido é fornecido. Aplicando pulsos de moduladores de via, este sistema pode ser usado para controlar externamente as oscilações de sinalização para a investigação funcional da sonorênese do mouse.

Uma abordagem microfluidics para a investigação funcional de oscilações de sinalização que regem a Somitogênese

Este artigo traça o ciclo de vida de um ressonador de membrana de nitreto de silício, desde o projeto até a fabricação e a caracterização.

Fabricação e caracterização de ressonadores de membrana de nitreto de silício High-Q

Um protocolo inovador de vedação para produzir lacres à base de silicone estanques para membranas de dispositivos de chip de Polidimetilsiloxano (PDMS). Destinado a laboratórios, configuração de modelos de chips e desenvolvimento de plataformas de chips em pequena escala. Demonstramos o protocolo usando membranas plásticas e de tecido nativo. Esse procedimento requer apenas PDMS, tolueno, mofo, membrana e uma câmara de vácuo.

Procedimento acessível de vedação de silicone chip-membrana para uma ligação flexível e confiável

Aprenda a detectar quebras em sítios frágeis comuns usando ChIP nativo de γH2A.X. Este método rápido e eficiente reduz o tempo de trabalho em pesquisas sobre instabilidade genômica em células cancerígenas.

Capturando quebras de local frágeis comuns por γH2A nativo. X ChIP

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