上海金畔生物科技有限公司代理AAT Bioquest荧光染料全线产品,欢迎访问AAT Bioquest荧光染料官网了解更多信息。
酶显色底物ADP-ribose-pNP
货号 | 11700 | 存储条件 | 在零下15度以下保存, 避免光照 | |
规格 | 1 mg | 价格 | 2604 | |
Ex (nm) | 405 | Em (nm) | ||
分子量 | 724.37 | 溶剂 | Water | |
产品详细介绍 |
简要概述
酶显色底物ADP-ribose-pNP是美国AAT Bioquest生产的用于过氧化物酶的试剂,ADP-核糖-pNP是用来评估多聚ADP-核糖聚合酶(PARP)活性的一种显色底物。在405nm处可检测释放的p-硝基酚吸光度。标准曲线的斜率通常用来转换产生的ADP-核糖-pNP比色底物的的吸光度。使用ADP-核糖-pNP作为显色底物,PARP-1具有最大Km和Vmax数值(分别为151 uM和1.30nmolmin-1mg-1),其次是Tankyrase-1(82uM和18pmolmin-1mg-l)和VPARP(46uM和2 pmolmin-1mg-l)。显色底物通常用来决定PARP-1,tankyrase-1和VPARP 的动力学参数,然后筛选PARP-1,tankyrase-1和VPARP小分子抑制剂。由于ADP-核糖-pNP连续实验分析比标准的非连续PARP试验更有优势,因此其可用于高通量筛选PARP-1,tankyrase-1和VPARP的活性以及相关抑制剂。金畔生物是AAT Bioquest的中国代理商,为您提供最优质的酶显色底物ADP-ribose-pNP。
适用仪器
光吸收酶标仪 | |
吸收: | 405nm |
推荐孔板: | 白色孔板 |
产品说明书
样品实验方案
溶液制备
1.储备溶液
所有未使用的储备溶液应分成一次性等分试样,并在制备后储存在-20°C。 避免反复冻融循环。
1.1 ADP-核糖-pNP原液:
在H2O中制备5-10mM的储备溶液。 注意:应立即使用原液。
2.工作溶液
ADP-核糖-pNP工作溶液:
通过用测定缓冲液(50mM Tris,10mM MgCl 2,pH 8.0)稀释储备溶液来制备0.25mM测定溶液。
样品操作步骤
1.将0.01mL /孔的样品溶液加入0.09mL /孔测定溶液中,使96孔透明板中的最终体积为0.1mL。
2.使用吸光度酶标仪在405nm处监测平板。
参考文献
A colorimetric substrate for poly(ADP-ribose) polymerase-1, VPARP, and tankyrase-1
Authors: Nottbohm AC, Dothager RS, Putt KS, Hoyt MT, Hergenrother PJ.
Journal: Angew Chem Int Ed Engl (2007): 2066
Pharmacological identification of P2X1, P2X4 and P2X7 nucleotide receptors in the smooth muscles of human umbilical cord and chorionic blood vessels
Authors: Valdecantos P, Briones R, Moya P, Germain A, Huidobro-Toro JP.
Journal: Placenta (2003): 17
Poly(ADP-ribose) polymerase as a key player in excitotoxicity and post-ischemic brain damage
Authors: Meli E, Pangallo M, Baronti R, Chiarugi A, Cozzi A, Pellegrini-Giampietro DE, Moroni F.
Journal: Toxicol Lett (2003): 153
Interactions of nucleotide cofactors with the Escherichia coli replication factor DnaC protein
Authors: Galletto R, Rajendran S, Bujalowski W.
Journal: Biochemistry (2000): 12959
Kinetic mechanism of nucleotide cofactor binding to Escherichia coli replicative helicase DnaB protein. stopped-flow kinetic studies using fluorescent, ribose-, and base-modified nucleotide analogues
Authors: Bujalowski W, Jezewska MJ.
Journal: Biochemistry (2000): 2106
Effects of caffeine and adenine nucleotides on Ca2+ release by the sarcoplasmic reticulum in saponin-permeabilized frog skeletal muscle fibres
Authors: Duke AM, Steele DS.
Journal: J Physiol (1998): 43
Adenine nucleotides regulate ADP-ribosylation of membrane-bound actin and actin-binding to membranes
Authors: Schroeder P, Just I, Aktories K.
Journal: Eur J Cell Biol (1994): 3
Trimeric G-proteins of the trans-Golgi network are involved in the formation of constitutive secretory vesicles and immature secretory granules
Authors: Barr FA, Leyte A, Mollner S, Pfeuffer T, Tooze SA, Huttner WB.
Journal: FEBS Lett (1991): 239
Inhibition and labeling of the Ca2(+)-ATPase from sarcoplasmic reticulum by periodate oxidized ATP
Authors: Mignaco J, Scofano HM, Barrabin H.
Journal: Biochim Biophys Acta (1990): 305