Name: chromatin immunoprecipitation assay for the identification of arabidopsis protein-dna interactions in vivo
Uploaded: 2016-01-14T16:00:00
Description: Watch this Scientific Journal Video about Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo at JoVE.com

The authors would like to acknowledge The Plant Cell for allowing the use of some data published in this journal to elaborate the representative results described here in Figure 4. This work was supported by the EU 7FP Marie Curie-Initial Training Network EpiTRAITS (Grant Agreement 316965), and by the Spanish Ministerio de Econom&#236;a y Competitividad (grants BIO2010-15589 and BIO2013-43098-R).

植物材料の1架橋（1時間） <ol><li>シロイヌナズナの実験に用いたライン成長（ - WT - 野生型変異体対を、および/またはタグを表すライン対目的のタンパク質のタグ付けされたバージョンを表す線は、任意のタンパク質を融合していない）大きなペトリ皿に12-18日間（150 mm）のMS寒天培地（1 L：1×ムラシゲ・スクーグ塩、10gのスクロース、0.5グラムのMES、pHを5.7（KOH）、1％寒天）。土壌やバー...

<ol>
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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simplified+formaldehyde+fixation+and+immunoprecipitation+technique+for+studying+protein-DNA+interactions.">A simplified formaldehyde fixation and immunoprecipitation technique for studying protein-DNA interactions.</a> Anal Biochem. 197 (1), 83-90 (1991).</li><li>Yu, C. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HISTONE+DEACETYLASE6+interacts+with+FLOWERING+LOCUS+D+and+regulates+flowering+in+Arabidopsis.">HISTONE DEACETYLASE6 interacts with FLOWERING LOCUS D and regulates flowering in Arabidopsis.</a> Plant Physiology. 156 (1), 173-184 (2011).</li><li>Zhou, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome-wide+profiling+of+histone+H3+lysine+9+acetylation+and+dimethylation+in+Arabidopsis+reveals+correlation+between+multiple+histone+marks+and+gene+expression.">Genome-wide profiling of histone H3 lysine 9 acetylation and dimethylation in Arabidopsis reveals correlation between multiple histone marks and gene expression.</a> Plant Mol Biol. 72 (6), 585-595 (2010).</li><li>Lafos, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dynamic+regulation+of+H3K27+trimethylation+during+Arabidopsis+differentiation.">Dynamic regulation of H3K27 trimethylation during Arabidopsis differentiation.</a> PLoS Genet. 7 (4), e1002040 (2011).</li><li>Barcala, M., Fenoll, C., Escobar, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Laser+microdissection+of+cells+and+isolation+of+high-quality+RNA+after+cryosectioning.">Laser microdissection of cells and isolation of high-quality RNA after cryosectioning.</a> Methods Mol Biol. 883, 87-95 (2012).</li><li>Carter, A. D., Bonyadi, R., Gifford, M. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+fluorescence-activated+cell+sorting+in+studying+plant+development+and+environmental+responses.">The use of fluorescence-activated cell sorting in studying plant development and environmental responses.</a> Int J Dev Biol. 57 (6-8), 545-552 (2013).</li><li>Deal, R. B., Henikoff, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+INTACT+method+for+cell+type-specific+gene+expression+and+chromatin+profiling+in+Arabidopsis+thaliana.">The INTACT method for cell type-specific gene expression and chromatin profiling in Arabidopsis thaliana.</a> Nat Protoc. 6 (1), 56-68 (2011).</li><li>Zeng, P. Y., Vakoc, C. R., Chen, Z. C., Blobel, G. A., Berger, S. 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ここで説明のChIPプロトコールは、タンパク質およびアラビドプシス植物の in vivo での特定のDNA配列間の相互作用を分析するための再現可能かつ強力な技術です。目的のタンパク質の結合部位の同定に成功し、関連する相互作用が実際に行われている植物器官や発達段階を適切に選択する必要があります。また、植物材料の適切な固定および超音波処理によりクロマチンの最適な...

近年中に、分子遺伝学的ツールおよびゲノムの広い範囲は、モデル種のシロイヌナズナで開発されてきました。この技術は、植物の開発が規制されている方法を理解する上で非常に進歩を促進してきました。モデルとしてシロイヌナズナを用いて研究し、発達過程の中で、開花時期の遺伝的制御が広範囲に解析されています。これらの研究は、植物は非常に正確にそのようなホルモンや植物の年齢などの内因性手がかりに、また季節1の自然なサイクルで開花時期を同期させるような日長や温度などの環境シグナルに応答して開花時期を調節することが示されています2。開花の時間変化を有するシロイヌナズナ変異体の単離および特性は、内因性および環境要因に応じて開花時期を調節する遺伝子の複雑なネットワークを発表における決定要因となっています。これらの遺伝子回路は、開花のスイッチとして機能するいくつかのマスター遺伝子のレベルで統合され、花芽の正確なタイミングは、開花促進し、花のインテグレータ遺伝子1,3の上流に働くの活動を抑制するのバランスに依存しています。 ゲノムアプローチの最近の使用によって支援開花開始の制御におけるその役割のために同定された遺伝子の機能解析は、開花時期の調節における転写制御の中心的な役割を明らかにしました。実際には、開花エンコード転写のマスター遺伝子の多くは、4因子 。また、クロマチンリモデリングタンパク質複合体の数は、開花のマスター遺伝子の発現に影響を​​与えます。それらの変化した開花時のために単離さシロイヌナズナ変異体の数は、クロマチン修飾因子の様々なコード化する遺伝子に変異を有することが判明しました。ヒストンで翻訳後修飾を導入異なるクロマチン改築Eテール、ヒストンバリアントによって標準的なヒストンはシロイヌナズナ5,6の開花の適切なレギュレーションのために必要であるDNAまたは交換する相対ヌクレオソームの位置を変更します。これらのクロマチンリモデリング活動のいくつかは、特定のヒストン残基におけるアセチル化やメチル化などの共有結合修飾の堆積または除去を触媒します。これらのヒストンマークは特に開花遺伝子の転写活性を調節するために、他のクロマチンリモデリング複合体、転写因子または転写機構の成分を補充する特殊なエフェクターによって認識されます。 クロマチン免疫沈降（チップ）は、目的のタンパク質のためのin vivoでの DNA結合部位の同定（ 図1）ことができます。この手順は、DNAに架橋タンパク質の特定の化学物質の能力を利用します。得られたDNA-タンパク質複合体は、次いで、特異的抗体AGAを用いて免疫沈降することができ選択したタンパク質に結合instの転写因子、クロマチン結合タンパク質、または特定の改変および異種エピトープ（一般に「タグ」と呼ばれます）。これらの免疫沈降物から精製されたDNAは、目的の特定の配列の濃縮のために評価するために定量的PCR（定量PCR）反応における鋳型として使用することができます。この方法では、転写因子の結合部位または特定の遺伝子でのヒストンマークの分布が7,8を確立することができます。また、大規模並列シーケンシングを可能にする次世代シーケンシング（NGS）と合わせ、チップ​​技術は、転写因子の結合部位のゲノムワイドな識別だけでなく、ヒストン修飾の除幕式エピ風景を可能にしました。さらに、遺伝子発現の同時分析は、転写調節因子の結合または特定のヒストンマークの堆積は転写activiと相関方法をモニタリングできます遺伝子9のTY。 シロイヌナズナにおけるチッププロトコルの使用は、転写調節因子の様々な開花とどのようにこれらの構造変化は、遺伝子発現に影響を与える5,6のマスター遺伝子のクロマチン組織に与える影響を評価することができました。以前の結果は 、短い日でシロイヌナズナ遺伝子座早期抽苔 （EBS）は 、この遺伝子のショーに開花し、開花FTのマスター遺伝子のアップレギュレーションの加速度を開花し、変異体のリプレッサーとして機能することを示しました。また、FTにおける機能喪失型変異は完全にFTが EBS変異体の早期の開花およびEBS 10開花このマスター遺伝子の抑制のために必要であることが必要であることを示す、EBS変異体植物の初期開花表現型を抑制、11は、EBS は、具体的には 、ヒストンH3のジ-に結合し、目にすることができるトリメチルPHDを含むタンパク質をコードしますFT 12のクロマチン媒介抑制におけるEBSの役割を示唆している電子のリジン残基4（H3K4me2 / 3）、。 ChIPのアプローチの使用は、シロイヌナズナPHD含有タンパク質EBS 10,11は、その発現12を抑制するために花のインテグレータ遺伝子FTの調節領域に結合することを実証しました。チップ技術の使用によって得られる追加のデータは、このタンパク質は、シロイヌナズナの開発の初期段階で開花のこのマスター遺伝子のクロマチンにおけるヒストンアセチル化、活性な転写の顕著な特徴の低レベルを維持するために必要とされることを示しました。一緒に、遺伝子および遺伝子発現データを有するこれらの観察は、このシロイヌナズナPHD含有タンパク質は、花の積分FT遺伝子12の発現を調節することにより、開花時間の微調整において中心的な役割を有していることを示しています。ここで紹介する作品は、ヒストンの分析のためだけでなく、他のためだけでなく、有益な最適化された方法を提供し、タンパク質、および効率の向上と低下し、実験時間に関連するクロマチン。また、この報告書は、チップ・プロトコルの使用は、クロマチンの修飾の変化や植物遺伝子の転写状態、およびシロイヌナズナの開花の発症に遺伝子発現制御への影響をどのようにこれらのクロマチン媒介機構との関係に新たな洞察を提供している様子を示しています。

<table border="0">
	<tbody>
		<tr>
			<td>MES</td>
			<td>Sigma</td>
			<td>M8250</td>
			<td></td>
		</tr>
		<tr>
			<td>MS (Murashige and Skoog Basal Salt Mixture)</td>
			<td>Sigma</td>
			<td>M5524</td>
			<td></td>
		</tr>
		<tr>
			<td>Formaldehyde 37%&#160;</td>
			<td>Sigma</td>
			<td>F8775-25</td>
			<td>Use under the fume hood</td>
		</tr>
		<tr>
			<td>Protease inhibitor mix cOmplete ULTRA Tablets, Mini, EDTA-free, EASYpack</td>
			<td>Roche</td>
			<td>5892791001</td>
			<td></td>
		</tr>
		<tr>
			<td>Bioruptor Standard sonication device</td>
			<td>Diagenode</td>
			<td>B01010002 (UCD200TO)</td>
			<td></td>
		</tr>
		<tr>
			<td>Glycine</td>
			<td>Sigma</td>
			<td>50046</td>
			<td></td>
		</tr>
		<tr>
			<td>QIAquick PCR Purification Kit</td>
			<td>Qiagen</td>
			<td>28104</td>
			<td></td>
		</tr>
		<tr>
			<td>Dynabeads&#174; magnetic beads coupled with protein A or protein G</td>
			<td>Life Technologies</td>
			<td>10003D/10001D</td>
			<td>Check manufacturer&#8217;s manual for antibody affinity</td>
		</tr>
		<tr>
			<td>Miracloth</td>
			<td>Merck Millipore</td>
			<td>475855</td>
			<td></td>
		</tr>
		<tr>
			<td>Triton&#8482; X-100 Surfact-Amps&#8482; Detergent Solution</td>
			<td>Life Technologies</td>
			<td>85112</td>
			<td></td>
		</tr>
		<tr>
			<td>(mouse, rat, rabbit&#8230;)-IgG</td>
			<td>Diagenode</td>
			<td>C15400001, C15420001, C15410206</td>
			<td></td>
		</tr>
		<tr>
			<td>Magnetic rack - DynaMag&#8482;-2</td>
			<td>Life Technologies</td>
			<td>12321D</td>
			<td></td>
		</tr>
		<tr>
			<td>H3K9/14ac polyclonal antibody - Premium&#160;</td>
			<td>Diagenode</td>
			<td>C15410200-10</td>
			<td></td>
		</tr>
		<tr>
			<td>Chelex&#174; 100 Resin</td>
			<td>Bio-Rad</td>
			<td>142-2832</td>
			<td></td>
		</tr>
		<tr>
			<td>Proteinase K</td>
			<td>Life Technologies</td>
			<td>17916</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Myc Tag Antibody, clone 4A6</td>
			<td>Millipore</td>
			<td>05-724</td>
			<td></td>
		</tr>
		<tr>
			<td>LightCycler&#174; 480 SYBR Green I Master</td>
			<td>Roche</td>
			<td>4707516001</td>
			<td></td>
		</tr>
	</tbody>
</table>

chromatin immunoprecipitation assay for the identification of arabidopsis protein-dna interactions in vivo

Intricate gene regulatory networks orchestrate biological processes and developmental transitions in plants. Selective transcriptional activation and silencing of genes mediate the response of plants to environmental signals and developmental cues. Therefore, insights into the mechanisms that control plant gene expression are essential to gain a deep understanding of how biological processes are regulated in plants. The chromatin immunoprecipitation (ChIP) technique described here is a procedure to identify the DNA-binding sites of proteins in genes or genomic regions of the model species Arabidopsis thaliana. The interactions with DNA of proteins of interest such as transcription factors, chromatin proteins or posttranslationally modified versions of histones can be efficiently analyzed with the ChIP protocol. This method is based on the fixation of protein-DNA interactions in vivo, random fragmentation of chromatin, immunoprecipitation of protein-DNA complexes with specific antibodies, and quantification of the DNA associated with the protein of interest by PCR techniques. The use of this methodology in Arabidopsis has contributed significantly to unveil transcriptional regulatory mechanisms that control a variety of plant biological processes. This approach allowed the identification of the binding sites of the Arabidopsis chromatin protein EBS to regulatory regions of the master gene of flowering FT. The impact of this protein in the accumulation of particular histone marks in the genomic region of FT was also revealed through ChIP analysis.

エイト主な手順は、成長し、植物材料、クロマチンの架橋、クロマチン分離、クロマチンの断片化、DNAとの間の複合体の選択的単離の収穫など、in vivoでのタンパク質-DNA相互作用の同定のために、このチッププロトコルに白羽することができます免疫沈降、タンパク質の消化、DNA精製、およびqPCR分析による目的のタンパク質（ 図1）。 ChIPプロトコルにおける重要なステッ?...

Watch this Scientific Journal Video about Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo at JoVE.com

Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo

クロマチン免疫沈降は、DNAは、インビボでのシロイヌナズナのタンパク質の結合部位を同定するための強力な技術です。この手順は、クロマチン架橋および断片化、目的のタンパク質に対する選択的な抗体を用いた免疫沈降、および結合したDNAの定量PCR分析を含みます。私たちは、シロイヌナズナ植物用に最適化され、単純なChIPアッセイを説明します。

シロイヌナズナタンパク質-DNA相互作用の同定のための免疫沈降アッセイクロマチン<em&gt;インビボ</em

Intricate gene regulatory networks orchestrate biological processes and developmental transitions in plants. Selective transcriptional activation and ...

The overall goal of this procedure is to unveil interactions between proteins and DNA in the chromatin of arabidopsis cells. This method can help answer key questions in the plant biology field, such as which genes are regulated by a given transcription factor, or what system modifications are associated with a transcriptional status of genes. The main advantage of this technique is that through the cross-linking step, it is possible to fix and capture changes in chromatin organization occurring during plant development.Though this method can provide a insight into protein interaction in arabidopsis thaliana, it also can be applied to different plant orders and adapted to other plant species. Demonstrating this procedure will be Dorota Komar and Alfonso Mouriz, two PhD students in our laboratory. After growing arabidopsis according to the text protocol, collect 1.5 grams of plant material in 50 milliliter tubes, and keep them on ice during cross-linking.Then, use a laboratory burner-heated needle to make little holes in the lids of the tubes. Add 40 milliliters of 1x pbs and 1.08 milliliters of formaldehyde to the tubes. Screw the lids on the tubes, and place the tubes in a desiccator.Vacuum infiltrate for 10 minutes, then carefully release the vacuum to prevent churning up the solution. After mixing the sample, repeat the vacuum and mixing steps. Upon infiltration, observe the plant material and confirm that it becomes slightly translucent and sinks to the bottom of the tube.Add 2.5 milliliters of two molar glycine to a final concentration of 0.125 molar and apply the vacuum for five minutes. After releasing the vacuum, discard the solution by inverting the tubes and allowing it to drip through the holes in the lids. Use pbs to rinse the plantlets twice, and rinse with water once.Then dry the plantlets on a paper towel before using liquid nitrogen to freeze. Per each immunoprecipitation, prepare 15 microliters of magnetic beads in a 1.5 milliliter microcentrifuge tube. Add one milliliter of chromatin immunoprecipitation, or chip dilution buffer, to the beads, mix by rotation, and place the 1.5 milliliter microcentrifuge tube in a magnetic rack.After allowing the beads to attach to the magnet for about a minute, use a pipetman to remove the supernatant before repeating the wash. Re-suspend the beads in 200 microliters of chip dilution buffer. For immunoprecipitation add the required amount of the specific antibody to one of the two tubes prepared for each plant blind.A key factor in successful chip experiment is the chosen antibody for the immunoprecipitation of the protein of interest. Antibodies raised against plant transcription factor can be useful for chip experiments, but the serum has to be thoroughly tested. For example, antibodies checked only in Western blot analysis are not necessarily valid for chip.As a negative control, add the same amount of non-specific IGG to the second tube. Incubate the tubes at four degrees Celsius overnight on a rotating wheel to allow the antibody to attach to the beads. Using a mortar and pestle, thoroughly grind the frozen plant material in liquid nitrogen until the powder becomes homogeneous and light green.Transfer the powder to a new 50 milliliter tube. Under a fume hood, add 30 milliliters of extraction buffer one, and mix well to soak the tissue powder. From this step on, keep the samples at four degrees Celsius at all times, and make sure that the tissue is completely thawed before moving forward.Clear the slurry by passing it through a filtration tissue with a pore size of 22 to 25 micrometers into a new 50 milliliter tube. Then centrifuge at 1, 000 times g and four degrees Celsius for 20 minutes. Gently decant the supernatant from the pellet, which should be about two milliliters in volume.With five milliliters of extraction buffer two, re-suspend the pellet. Then centrifuge at 1, 000 times g and four degrees Celsius for 10 minutes. After decanting the supernatant, re-suspend the pellet in 300 microliters of extraction buffer three.In a separate tube, add 600 microliters of extraction buffer three and then carefully layer the re-suspended pellet on top of the clean buffer. Centrifuge the tube for one hour at 16, 000 times g at four degrees Celsius, followed by aspiration of the supernatant with a pipette. Next, use 300 microliters of sonication buffer to slowly re-suspend the nuclear pellet, avoiding bubble formation, which may affect sonication efficiency.Carefully transfer the suspension to a clean, 1.5 milliliter tube. Sonicate the suspension to lyse the nuclei and randomly share the chromatin into 200 to 600 base pair fragments. Verify the DNA fragment sizes on an agarose gel, then spin the solution at 12, 000 times g and four degrees Celsius for 10 minutes.Transfer the supernatant to a fresh 1.5 milliliter tube, and discard the pellet. Aliquot one-tenth of the supernatant into a new tube, and label it as input'before freezing at minus 20 degrees Celsius. Finally, make a ten-fold dilution of the chromatin in chip dilution buffer to achieve a final SDS concentration of 0.1 percent.Samples can then be frozen and stored at minus 20 degrees Celsius for several weeks. After incubating the beads with antibodies overnight, use one milliliter of chip dilution buffer to wash the beads three times, then re-suspend the beads in one milliliter of the diluted chromatin, and incubate the tube overnight on a rotating wheel at four degrees Celsius. The following day, place the beads on the magnetic rack at four degrees Celsius, and after removing the solution, use one milliliter of low-salt washing buffer to wash the beads twice, then use high-salt washing buffer to wash the beads once.After using one milliliter of lithium chloride washing buffer to wash the beads once, use one milliliter of TE buffer to wash the beads twice. Aspirate to thoroughly remove the TE wash buffer. Once the input samples are thawed, transfer five microliters of each into new 1.5 milliliter tubes, and moving forward, treat similarly to the chip samples.Reverse the cross-linking by adding 200 microliters of five percent chelating ion exchange resin and incubate at 95 degrees Celsius for 10 minutes, shaking every two to three minutes. Spin down at 16, 000 times g for 30 seconds, and carefully transfer the supernatant into new microfuge tubes. Add two microliters of 10 milligrams per milliliter protonase K, and incubate at 37 degrees Celsius for 30 minutes to digest proteins and release DNA.To stop the reaction, incubate at 95 degrees Celsius for 10 minutes, Quickly spin at 16, 000 times g for 30 seconds, and transfer the supernatant to a new tube. After cleaning the DNA according to the text protocol, dilute one to ten with water, and use five microliters per reaction to measure the abundance of binding sites by qPCR. Analyze the data according to the text protocol.Aribidopsis leaves are covered in wax layers and trichomes favor the formation of air bubbles, both of which make it difficult for cross-linking reagents to penetrate the plant tissue. Shown here are cross-linked seedling samples that were vacuum cross-linked to increase fixation efficiency. This figure demonstrates that an increased number of sonication cycles progressively reduces the average size of the DNA in chip samples, reaching the optimum enrichment in the range of 200 to 600 base pairs after 20 to 30 cycles, as demonstrated here.As seen in this figure, four regions important for the transcriptional regulation of the FT locus, a master gene of flowering, have been chosen for the analysis. Chip results demonstrate that the EBS protein combined one of the four regions tested, a regulatory sequence close to the transcriptional start site of FT.Finally, chip assays performed using an antibody directed against histone h3 acetylated in lysines nine and 14, which is correlated with transcriptionally active chromatin, revealed higher abundance of H3K9/14ac in mutant DBS versus wild type plants in all four genomic fragments of FT tested, consistent with the observed up-regulation of this master gene of flowering in this mutant. Once mastered, this technique can be done in two days if it is performed properly.Following this procedure, together with other measures like gene expression analysis, can contribute to answering other questions, like:What is the biological role of the binding of the protein to DNA? Is it at transcriptional activation or repression? And Which histone modifications are associated?After it's development, this technique paved the way for researchers in the field of plant epigenetics to explore chromatin states in our reductive cells. And this can be extended to other plant species. After watching this video, you should have a good understanding of how to analyse the binding of your aribidopsis protein of interest to DNA in vivo.Don't forget that working with 2-methoxyethanol, formaldehyde or sonicator reagents, pathogens and stem mutations can be extremely hazardous, and precautions such as protective clothes, gloves or hearing protection should be always taken while using this procedure.

Research

JoVE Journal

Biology

Chromatin Immunoprecipitation Assay for the Identification of Arabidopsis Protein-DNA Interactions In Vivo

Chromatin Immunoprecipitation from Human Embryonic Stem Cells

ヒト胚性幹細胞からクロマチン免疫沈降

The functional and structural complexity of the myriad of cells in metazoan organisms arises from a small number of stem cells. Stem cells are ...

生物学

Chromatin Immunoprecipitation (ChIP) to Assay Dynamic Histone Modification in Activated Gene Expression in Human Cells

Chromatin Immunoprecipitation ChIP to Assay Dynamic Histone Modification in Activated Gene Expression in Human Cells

ヒト細胞において活性化遺伝子発現のアッセイのダイナミックヒストン修飾のクロマチン免疫沈降（ChIP）

In response to a variety of extracellular ligands, the STAT (signal transducer and activator of transcription) transcription factors are rapidly recruited ...

Chromatin Immunoprecipitation Assay for Tissue-specific Genes using Early-stage Mouse Embryos

初期段階のマウス胚を用いて組織特異的遺伝子のためのクロマチン免疫沈降アッセイ

Chromatin immunoprecipitation (ChIP) is a powerful tool to identify protein:chromatin interactions that occur in the context of living cells 1-3. This ...

Chromatin Immunoprecipitation (ChIP) using Drosophila tissue

Chromatin Immunoprecipitation ChIP using Drosophila tissue

を使用してクロマチン免疫沈降（ChIP）ショウジョウバエ組織

Epigenetics remains a rapidly developing field that studies how the chromatin state contributes to differential gene expression in distinct cell types at ...

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

A Protocol for the Identification of Protein-protein Interactions Based on 15N Metabolic Labeling, Immunoprecipitation, Quantitative Mass Spectrometry and Affinity Modulation

に基づいて、タンパク質 - タンパク質相互作用の同定のためのプロトコル<sup&gt; 15</sup&gt; Nの代謝標識、免疫沈降法、定量的質量分析法と親和性調節

Protein-protein interactions are fundamental for many biological processes in the cell. Therefore, their characterization plays an important role in ...

Efficient Chromatin Immunoprecipitation using Limiting Amounts of Biomass

バイオマスの制限額を使用して効率的なクロマチン免疫沈降

Chromatin immunoprecipitation (ChIP) is a  widely-used method for determining the interactions of different proteins with  DNA in chromatin of living ...

Combining Single-molecule Manipulation and Imaging for the Study of Protein-DNA Interactions

タンパク質-DNA相互作用の研究のための単一分子マニピュレーションとイメージングを組み合わせる

The paper describes the combination of optical tweezers and single molecule fluorescence detection for the study of protein-DNA interaction. The method ...