拓扑根/人拓扑异构酶I(Top1)/TG2005H-RC3/3000单位(300 u/ul)

价格
¥17842.50
货号:TG2005H-RC3
浏览量:127
品牌:TopoGEN
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商品描述

HumanTopoIProductBackground
TopoGENnowoffersahighpurityandcatalyticallyactiveTop1Enzymeoverexpressedinthebaculovirussystemandpurifiedtosinglebandhomogeneity. ThesepreparationsarecheaperthannativetopoImadefromhumantissueandofferexcellentpurityandhighspecificactivity. Inaddition,the100kDaformof recombinanttopoIisstableandislesspronetoproteolysis,relativetothenativeproteinfromhumancells.

QualityControlTests:

  • 1
  • AtestfornucleasecontaxaminationwascarriedoutbyassayingfortheformationoflinearKDNAandlinearplasmidDNA. Incubationsof1µgofcatenatedKDNAorsupercoiledpUC19DNA(4hrs.at37°Cinthepresenceof10mMMgCl2)wereperformed. LinearDNAorbreakdownproductswerenotgeneratedundertheseconditions.
  • 2
  • AcheckforcrosscontaminationwithtopoIIwasnegative. TherewasnodecatenationofkDNAintopoIIreactionconditions.
  • 3
  • ThefinalfractionoftopoisomeraseIisacolumnpoolandisinthefollowingbuffer:20mMNaH2PO4(pH7.4),300mMNaCl,50ug/mlBSA,50%glycerol,50mMImidazole*. ThefinalfractionwasanalyzedbySDS-PAGEandshowntocontainasingle,predominantbandof100kDa.
    *Actualconcentrationcanrangefrom25mM-100mM,accordingtohowstockenzymewasdilutedinstoragebuffer. 
 

ActivityAssays:
Relaxationassayswerecarriedoutinafinalvolumeof25µlinrecombinanttopoIreactionbuffer(10Xreactionbuffer,suppliedwiththisproductis:100mMTris-Cl,pH7.9,1.5MNaCl,1%BSA,1mMSpermidine,50%glycerol).  SupercoiledplasmidDNAwasincludedat0.25µg/reaction. Reactionsterminatedwith5µl(per20µlreactionvolume)ofstopbuffer(5%sarkosyl,0.0025%bromophenolblue,25%glycerol).Reactionproductswereanalyzedona1%nativeagarosegel. Undertheseconditions,relaxationactivityiseasilydetectable(typicallywesee10-40unitsofactivityperul). Atypicaltitrationisshownbelow:
 

Topoisomerase

  • ReviewsandCitations:
  • YoshizakiA,YanabaK,OgawaA,AsanoY,KadonoT,SatoS:ImmunizationWithDNATopoisomeraseIandFreund'sCompleteAdjuvantInducesSkinandLungFibrosisandAutoimmunityviaInterleukin-6Signaling. ArthritisandRheumatism2011,63:3575-3585. doi:10.1002/art.30539.
  • ZhangL,SullivanP,SuyamaJ,et.al:Epidermalgrowthfactor-inducedheparanasenucleolarlocalizationaugmentsDNAtopoisomeraseIactivityinbrainmetastaticbreastcancer. MolecularCancerResearch2010,8:278-290.doi:10.1158/1541-7786.MCR-09-0375

IncludedMaterials:
10Xassaybufferanddilutionbufferisincluded. ThesemaybepurchasedseparatelyasTGS10XTopoIAssayBuffer(TG4095).

TopoGEN人类拓扑异构酶IIα单克隆抗体SKU尺寸价钱数量TG2010-150伙$ 285.00                  1个                  2                  3                  4                  5                  6                  7                  8                  9                  11                   添加到购物车人类拓扑异构酶IIα单克隆抗体:免疫印迹和免疫组化试剂制备了针对源自p170 kDa人Topo IIa C端的寡肽的单克隆抗体。然后在含有相同肽的柱子上亲和纯化抗体。亲和纯化的抗体可用于探测Western印迹,前提是包括对照以清楚地标记170 kDa topo II的位置。原因是在常规提取和操作过程中人topo II很容易降解或被蛋白水解。结果,可以检测子带。一些实验室已经报道了SDS-PAGE模式的异质性(Shelton等,1983; Halligan等,1985; Muller等,1988),最近还报道了粟酒裂殖酵母拓扑异构酶II的蛋白水解产物(Shiozaki和Yanagida,1991年)。因此,如果存在广泛的蛋白水解,抗体可能看起来是非特异性的。TG2011-3),其对应于完整的170kDa拓扑异构酶II。报道了编码汉拓扑异构酶II的第二个基因(拓扑II-β或180kDa形式)(Chung等,1989; Tan等,1992)。该抗体与topo II的170 kDa亚型特异性反应。单克隆抗体单位定义:抗体浓度以蛋白质印迹单位为单位;一个单位对应于制备常规蛋白质印迹稀释探针所需的抗体的1:1000稀释度。因此,10个单位的抗体将制成10 ml稀释的探针(直接添加到单个印迹中)。