當(dāng)與酶活性重塑復(fù)合物如SMARCA4、SMARCA2和ACF（如EpiCypher貨號(hào)15-1014、15-1015、15-1013）配對(duì)時(shí)，該產(chǎn)品提供核小體重塑的高通量熒光讀數(shù)。EpiDyne?-FRET單核小體由在大腸桿菌中表達(dá)的重組人組蛋白（組蛋白H2A-Cy5、H2B、H3.2和H4各兩種；accession numbers:：H2A-P4908；H2B-O60814；H3.2-Q71DI3；H4-P62805）組裝而成，該組蛋白由212個(gè)DNA堿基對(duì)纏繞，并攜帶5'Cy3、Widom 601核小體定位元件和重塑時(shí)暴露的DpnII限制性內(nèi)切酶基序（GATC）[1]。H2A在殘基120位處具有Thr-Cys取代，其中Cy5是共軛的。H3.2在殘基110位處具有Cys-to-Ala取代。
[1] Lowary & Widom J Mol. Biol. (1998) PMID: 9514715.
產(chǎn)品詳情
保存溫度:Stable for six (6) months at -80°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.
運(yùn)輸溫度:Dry ice.
產(chǎn)品形式:Purified recombinant mononucleosomes, (21.8 μg protein weight, 50 μg DNA+protein) in 44.2 μL of 10 mM Tris-HCl pH 7.5, 25 mM NaCl, 1 mM EDTA, 2 mM DTT, 20% glycerol. MW = 240,650 Da. Molarity = 4.7 μM.
驗(yàn)證數(shù)據(jù)
 | Figure 1: DNA Gel Data EpiDyne-FRET substrate resolved via native PAGE gel and? stained with ethidium?bromide to visualize DNA. Lane 1: Free? DNA (100 ng). Lane 2: Intact EpiDyne-FRET?nucleosomes? (400 ng). |
 | Figure 2: Protein Gel Data Coomassie stained PAGE gel of proteins in EpiDyne-FRET? substrate (1 μg)?demonstrates the purity of histones in the? preparation. Sizes of molecular?weight markers and positions of the core histones (H2A-Cy5, H2B, H3.2, and H4)?are? indicated. |
 |
Figure 3: Nucleosome Remodeling Data ACF1/ATP-dependent nucleosome remodeling reaction. EpiDyne-FRET? nucleosomes (15 nM) were?incubated with ACF1 chromatin remodeler ?(EpiCypher 15-1013; indicated concentrations) in the presence?or absence? of 1 mM ATP. Upon the addition of ATP, reactions were immediately read? in an Envision Multilabelplate reader (PerkinElmer). Data are presented? as the mean of the Cy3/Cy5 ratio (N=2). |
訂購(gòu)詳情
貨號(hào) | 產(chǎn)品名稱 | 規(guī)格 |
16-4201? ? | EpiDyne-FRET Nucleosome Remodeling Assay Substrate? ? | 50 μg? ? |
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