
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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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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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?
不同的酶Km值不同,同一种酶与不同底物反应Km值也不同,Km值可近似的反应酶与底物的亲和力大小:Km值大,表明亲和力小;Km值小,表明亲合力大。
Km最小的那个底物,就是酶的最适底物。
我是新手,来到丁香园看到有这么多的热心人,感到很高兴!!我也有很多问题需要大家的帮助!!实验已经作了一年了可是毫无进展,心里很是着急!!
我的课题是以外消旋的苯基乙二醇为底物,用假丝酵母催化生成手性纯的S-型苯基乙二醇,由于是老课题,所以目标是提高转化反应的底物浓度和菌体的使用批次(目前菌体使用一批后便不能再使用)。
我曾试过很多种方法,但均效果不大!我试着在转化过程中添加醛类,酮类,醇类作为辅助底物,增加菌体的使用批次。可效果不好,特别是添加了醇类后还有的起了反作用,因为我的这个转化过程中涉及到NAD和NADPH的再生。(其转化过程是酵母先催化将外消旋的苯基乙二醇变为酮,再将酮还原为醇,经过这一过程就将外消旋的苯基乙二醇变为手性纯的S-型了)
我还试过用固定化的方法,海藻酸钙包埋法,可是这样底物浓度就更低了!!
我现在不知道下一步该如何做了,很着急,请大家帮帮忙。谢谢了!!谢谢

