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
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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
如果能分解,那小肠液中的消化酶如何大量共存,如果不能
那它如何识别其他蛋白质物质是不是消化酶?
反应体系如下:
plasmid10ul
10xKbuffer5ul
KpnI1ul
BamHI1ul注意千万不可多加总酶量必须<4%
ddH20upto50ul
总体积改变加酶量按比例改变.
明白了么,少年?
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
暂无品牌问答