DESCRIPTION:
Human recombinant BRD4, (residues 1-744), like other human members of the BET family of chromatin-binding proteins (BRD2, BRD3, BRDT), comprises two bromodomains, protein modules that bind e-N-acetyllysine residues. The ubiquitously expressed BRD4 functions as a transcriptional regulator with roles in cell cycle progression and has recently been shown to be an atypical kinase that can phosphorylate RNA Pol II. Recent structural studies have shown that BRD4-1, like the bromodomain-1 of fellow BET family protein BRDT, can bind simultaneously to two acetyllysine residues with appropriate spacing and sequence context, for example a histone H4 peptide acetylated at lysines 5 and 8 (H4K5AcK8Ac). Chromosomal translocations that produce BRD4-NUT fusion proteins are implicated in causation of a rare and aggressive cancer, NUT midline carcinoma. Selective inhibitors of BRD4/BET family bromodomains are showing promise as possible therapeutic agents for cancer and inflammation.
ACCESSION #: NM_058243
Uniprot Link
INCLUDES AMINO ACIDS: 1-744
TAG(S):N-terminal StrepII-tag and C-terminal His-tag
MW: 85.7 kDa
EXPRESSION SYSTEM: Sf9 insect cells
SUPPLIED AS: Solution of purified recombinant protein in 50 mM Tris/HCl, pH 8, 500 mM NaCl, 10% glycerol (v/v)
STORAGE:-70°C. Thaw quickly and store on ice before use. The remaining, unused, undiluted protein should be snap frozen, for example in a dry/ice ethanol bath or liquid nitrogen. Minimize freeze/thaws if possible, but very low volume aliquots (<5 µl) or storage of diluted enzyme is not recommended.
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Substantial Discounts Available on Bulk Quantities
eactionBIOLOGy公司成立于2001年,是专门从事早期药物发现化验服务的公司,提供广泛的服务,包括用于激酶和表观遗传生物化学以及基于细胞的高通量筛选(HTS)和选择性化合物分析的稳定测定技术。我们还提供相关的HTS检测试剂,包括高度纯化和特征化的表观遗传蛋白。平均每年进行数百万次HTS和性能分析。在全球有超过750家公司(包括制药,生物技术,学术和非营利实验室)购买我们的产品。
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ReactionBiology详细产品列表:货号产品价格(美元)HMT-11-376ASH1L(His)Methyltransferase325DMT-21-124DNMT1450DMT-21-125DNMT3aMethyltransferase450DMT-21-126 DNMT3b450HMT-11-101DOT1LMethyltransferase395HMT-25-115 EZH1Complex450HMT-25-114EZH2Complex799HMT-25-173EZH2-Y641FComplex799HMT-11-102G9aMethyltransferase(GST)(aa786-1210)299HMT-11-245G9aMethyltransferase(His)(aa913-1193)299HMT-12-104G9a/GLPComplexMethyltransferase325HMT-11-103GLPMethyltransferaseProtein299HMT-15-105MLL1Complex499HMT-15-106 MLL2Complex499HMT-15-107MLL3Complex499HMT-15-108 MLL4Complex499MT-11-359NNMT(GST)(NicotinamideN-Methyltransferase)299MT-11-358NNMT(His)(NicotinamideN-Methyltransferase)299HMT-11-319NRMT1(GST)325HMT-21-139NSD1Methyltransferase450HMT-21-1380NSD2(His)Methyltransferase450HMT-21-122 NSD2Methyltransferase550HMT-11-377NSD2SET(His)325HMT-21-159NSD2-E1099KMethyltransferase599HMT-11-378NSD2-E1099K-SET(His)325HMT-21-181NSD2-T1150AMethyltransferase450HMT-11-379NSD2-T1150A-SET(His)325HMT-11-132 NSD3Methyltransferase325HMT-21-348PRDM2Methyltransferase325HMT-21-152PRDM9Methyltransferase450HMT-11-119PRMT1Methyltransferase325HMT-11-113PRMT3Methyltransferase325HMT-11-120PRMT4Methyltransferase399HMT-21-172PRMT5Methyltransferase450HMT-22-434PRMT5(C449S)/MEP50799HMT-22-148PRMT5/MEP50Complex799HMT-11-121PRMT6Methyltransferase(GST)325HMT-21-380PRMT6Methyltransferase(His)325HMT-21-382PRMT7(His)450HMT-11-135PRMT8Methyltransferase325MT-11-356s-COMT(His)(SolubleCatechol-O-Methyltransferase)299MT-11-357s-COMT(V108M)(His)(SolubleCatechol-O-Methyltransferase)299HMT-15-116 SET1AComplex(5-subunits)499HMT-15-117SET1BComplex(5-subunits)499HMT-11-133SET7/9Methyltransferase299HMT-11-476SET8(GST)299HMT-11-118 SET8Methyltransferase299HMT-11-128 SETD2(GST)Methyltransferase299HMT-11-129SETD2(His)Methyltransferase299HMT-11-110SMYD2Methyltransferase325HMT-11-111SUV39H1Methyltransferase299HMT-11-418SUV39H2(GST)Methyltransferase299HMT-11-112SUV39H2(His)Methyltransferase299HMT-21-149SUV420H1-tv1Methyltransferase450HMT-21-150SUV420H1-tv2Methyltransferase450HMT-21-349SUV420H2Methyltransferase325HMT-14-109WRAD2375HMT-14-438(H3.3-H4)2Tetramer325HMT-35-435CoreHistones(Chicken)250HMT-11-184Fibrillarin(GST)325HMT-11-183Fibrillarin(His)325HMT-11-137GST-GAR299HMT-12-316H2A/H2Bdimers325HMT-11-180HistoneH1.050HMT-11-146HistoneH2A50HMT-11-147HistoneH2B50HMT-11-134 HistoneH3.350HMT-35-179Nucleosomes(ChickenMono/Di)280HMT-35-182Nucleosomes(ChickenOligo/PolyH5Enriched)280
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调节至最适合温度(一般是37度),合适的酸碱度时,酶的活性最高
ELISA实验,操作的时候,加入底物,溶液变成蓝色,孵育一段时间后,加入终止液,变成黄色,测吸光值。然后,当天忘记扔板子了,第二天想到去扔的时候,所有孔都变成无色透明的了!!!为什么会褪色???是试剂有问题?还是因为室温太高环境影响?如果是试剂问题那我之前做的结果还能信么?
1,底物DNA上没有该限制酶的识别、切断位点。特别是一些经过重组等处理的DNA,碱基易发生缺失、变化等。
2,限制酶识别位点上的A或C被甲基化。部分限制酶对识别位点中的碱基是否被甲基化比较敏感,从而无法切断该位点。
3,底物不纯。如果底物DNA中有限制酶阻害物质,回影响限制酶的酶切作用。在此种情况下,底物DNA须重新进行精制。
4,限制酶的识别、切断位点在底物DNA的高级构造中所处的位置,对酶切反应也有一定的影响,例如,限制酶NaeI在切断pBR322DNA时,就有着非常难以切断的部位。
5,限制性内切酶本身无活性或低活性