
LifeSensors’sdiubiquitinsubstratesrepresentanewclassofsubstratesforthecontinuousfluorescentmeasurementoftrueisopeptidaseactivity.TheC-terminusofwildtypeubiquitinisconjugatedviaanisopeptidebondtolysine48(K48)ofasecondubiquitinmoleculewiththeresultantdiubiquitinforminganinternallyquenchedfluorescentFRETpair(IQF).Eachubiquitinislabeledwithasinglemoleculeofeitherafluorescentreporter(i.e.TAMRA)orahighlyefficientquenchingdye.CleavageoftheIQFDiUbbydeubiquitylasesleadstoseparationofthefluorophorefromquencherandsubsequentincreaseinobservedfluorescence.
ThereisaremarkabledifferencebetweenK48-andK63-linkedubiquitinmoleculesintheirthree-dimensionalstructure,whichaccountsfortheirindividualfunctions.Beyonddifferentiallinkages(e.g.K48,K63,K11),LifeSensorshascreatedsubpanelsofIQFdiubiquitinsubstrateswithineachlinkage. BecauseeachDUBislikelytorecognizeandcleavesubstrateswithuniquestericconsiderations,thesesubpanelsvaryinlocationofreporterfluorophoreandquencher. WerecommendthateachDUBbeempiricallyevaluatedagainstapanelofIQFDiUbsubstratestoselectthetheoptimalfluorophore/quencherpairing.
IQFDiUbsubstratepanels
InordertoefficientlydeterminetheoptimalsubstrateforanyDUB,orrapidlyassessapanelofvariousDUBs,LifeSensorsofferscomprehensivepanelsforeitherK48 (DiUb48panelcatno.DU0101) orK63linkeddiubiquitins (DiUb63PanelDU0102).Eachpanel containallsixvariantsoftherespectivelinkagealongwithacontrolenzyme(USP2core).Inadditionthereisalsoafull(K48/K63) panel (DU0201) thatcontainsall12variants.
Needbulkquantities?Wewouldbehappytoworkoutacustompricethatfitsyourbudget.Pleaseemail info@lifesensors.com formoreinformation.
ThisproductiscoveredbyUSpatent#8,518,660. Bypurchasingthisproduct,thepurchaseragreestocomplywiththetermsofour LimitedUseLabelLicense.
Info
Amount | 50µg |
State | Liquid |
Concentration | 20µM |
Buffer | 50mMSodiumMES,pH6.0 |
Variant | 2 |
Linkage | K48 |
Molecularweight | 18,548.4+/-18Da |
Fluorescentreadout | TAMRA |
Storage | IQFDiUbsarestablefor4-6monthsat4ºC.Longterm(>6months)storageat-80ºCisrecommended.Avoidrepeatedfreeze/thawcycles. |
Applications
- Kineticcharacterizationofdeubiquitylases(DUBs)
- DemonstrationofnovelDUBactivityand/orlinkagespecificity
- MonitoringofheterologousDUBproduction/purification
- Highthroughputscreening(HTS)forantagoNISTsoragonistsof deubiquitylaseactivity
- Additionalmaterialsrequired(forfurtherdetails,pleaseseemanual):
- 384-(GreinerBioOne781209) or96-well(655076)blackplatesorequivalent
- Fluorescentplatereader
- FiltersormonochromatorsformonitoringfluorescenceofTAMRA(Ex540nm,Em580nm)
- Dichroicmirrorwitha550-570nmcutoffisrecommendedforoptimalsignal-to-background
Benefits
- MoreBIOLOGicallyrelevantsubstratecomparedtotrADItionalmono-ubiquitinderivatives.
- FluorophoresareNOTattheN-orC-terminus,modificationsknowntoperturbubiquitinstructure.
- AvarietyoflinkagesandfluorophorepositioningavailableforoptimalactivityofyourDUBofinterest.
- Validatedinboth96-welland384-wellformat.
Documents
ebiomall.com






>
>
>
>
>
>
>
>
>
>
>
>
请教各位老师,
文献中检测细胞的活性有的用“NADPHdehydrogenase(NADPH脱氢酶)”有的用“NADPHdiaphorase(NADPH黄递酶)”
它们是同一种酶吗?它们的功能是什么?检测它们的活性是否能够评估细胞的活性。
不胜感激
有谁用碧云天的过氧化氢酶检测试剂盒,在试剂盒的准备工作中,过氧化氢的实际浓度=22.94*A240,我测出来的吸光度是3.3左右,那么乘以22.94就等于76多点,再乘以之前的稀释倍数,大约就是7600mM左右,这样跟说明书中说道的1M相差太多,感觉不对啊,稀释的肯定是没有错的,但是不知道哪里出了错,怀疑说明书就有问题呢,有人做过这个实验吗?能不能准确测出过氧化氢浓度吗?有测过的人帮忙指点一下啊,不知道应该怎么做
胶原酶按其存在的方式不同可分为人体内源性胶原酶和药用胶原酶两种。人体内源性胶原酶是指人体内部本身所具有的胶原酶,如牙龈、触膜等上皮组织和关节滑膜、椎间盘内都不同程度的存在着这种胶原酶,它在体内胶原蛋白的分解过程中发挥着不可或缺的作用。药用胶原酶是指利用生物制药的高科技手段从溶组织梭状芽孢杆菌的发酵液中提取、纯化并精制而得的白色或类白色无菌冻干粉针生物制剂。
(2)不同点:①影响因素:变构调节是由细胞内变构效应剂浓度的改变而影响酶的活性;化学修饰调节是激素等信息分子通过酶的作用而引起共价修饰。②酶分子改变:变构效应剂通过非共价键与酶的调节亚基或调节部位可逆结合,引起酶分子构像改变,常表现为变构酶亚基的聚合或解聚;化学修饰调节是酶蛋白的某些基团在其他酶的催化下发生共价修饰而改变酶活性。③特点及生理意义:变构调节的动力学特征为S型曲线,在反馈调节中可防止产物堆积和能源的浪费;化学修饰调节耗能少,作用快,有放大效应,是经济有效的调节方式。

