Epicypher热销产品——EpiDyne-FRET Nucleosome Remodeling Assay Substrate

2023-10-26

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当与酶活性重塑复合物如SMARCA4、SMARCA2和ACF（如EpiCypher货号15-1014、15-1015、15-1013）配对时，该产品提供核小体重塑的高通量荧光读数。EpiDyne®-FRET单核小体由在大肠杆菌中表达的重组人组蛋白（组蛋白H2A-Cy5、H2B、H3.2和H4各两种；accession numbers:：H2A-P4908；H2B-O60814；H3.2-Q71DI3；H4-P62805）组装而成，该组蛋白由212个DNA碱基对缠绕，并携带5'Cy3、Widom 601核小体定位元件和重塑时暴露的DpnII限制性内切酶基序（GATC）[1]。H2A在残基120位处具有Thr-Cys取代，其中Cy5是共轭的。H3.2在残基110位处具有Cys-to-Ala取代。

[1] Lowary & Widom J Mol. Biol. (1998) PMID: 9514715.

产品详情

保存温度:Stable for six (6) months at -80°C from date of receipt. For best results, aliquot and avoid multiple freeze/thaws.

运输温度:Dry ice.

产品形式: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.

验证数据

	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).