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 | 25µL |
| State | Liquid |
| Concentration | 20µM |
| Buffer | 50mMSodiumMES,pH6.0 |
| Variant | 2 |
| Linkage | K63 |
| 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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1、洗碗用的洗洁精,含脂肪酶,能快速水解油脂,还可能含其他酶成分
2、洗头用的洗发水,含脂肪酶,能快速水解油脂,还可能含其他酶成分
3、洗澡用的沐浴露,含脂肪酶,能快速水解油脂,还可能含其他酶成分
4、洗衣用的洗衣粉或洗衣液,含有多种酶,比如脂肪酶、蛋白酶等
除了以上用的频繁外,还有一些可能用到的,
比如某些促进消化的药片,含有脂肪酶、蛋白酶、淀粉酶等
厨房用的嫩肉粉,含有蛋白酶等
那糖酵解中依赖NAD的酶不是要在线粒体内反应?
因为溶蛋白酶是蛋白质本质的酶
食物酶:
天然存在於所有的生食物中。他们是消化酶的外部来源。食物酶在烹饪和处理过程中很容易被破坏。
一些富含酶的食物:
青木瓜内含丰富的木瓜酵素、木瓜蛋白酶、凝乳蛋白酶、胡萝卜素。
绿豆富含维生素B族、葡萄糖、蛋白质、淀粉酶、氧化酶。
生的蔬菜水果坚果种子富含各种酶,如菠菜、海藻等。
胡萝卜富含维生素C分解酶,萝卜、香瓜、菜花富含过氧化物酶,南瓜含维生素C分解酶,蕨菜含有维生素B1分解酶,菠萝和猕猴桃富含蛋白酶,无花果富含淀粉酶和蛋白酶,纳豆菌中富含淀粉酶、纤维酶。
人体内存在大量酶,结构复杂,种类繁多,到目前为止,已发现3000种以上(即多样性).如米饭在口腔内咀嚼时,咀嚼时间越长,甜味越明显,是由于米饭中的淀粉在口腔分泌出的唾液淀粉酶的作用下,水解成麦芽糖的缘故.因此,吃饭时多咀嚼可以让食物与唾液充分混合,有利于消化.此外人体内还有胃蛋白酶,胰蛋白酶等多种水解酶.人体从食物中摄取的蛋白质,必须在胃蛋白酶等作用下,水解成氨基酸,然后再在其它酶的作用下,选择人体所需的20多种氨基酸,按照一定的顺序重新结合成人体所需的各种蛋白质,这其中发生了许多复杂的化学反应.可以这样说,没有酶就没有生物的新陈代谢,也就没有自然界中形形色色、丰富多彩的生物界.
蛋白酶分布广泛,主要存在于人和动物消化道中,而消化道通过口与肛门与外界相通,属于外环境或外界环境
(溶菌酶、凝血酶原等)分布在内环境
(消化)酶分布在消化道

