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 ethidiumbromide to visualize dna. lane 1: free
dna (100 ng). lane 2: intact epidyne-fretnucleosomes
(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 molecularweight 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) wereincubated with acf1 chromatin remodeler
(epicypher 15-1013; indicated concentrations) in the presenceor 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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