ProductHighlight

• Configuration-specificmonoclonalantibodythatrecognizesCdc42-GTP,butnotCdc42-GDP
• TheonlyavailableCdc42activationassaykitdirectlymeasuresthelevelofCdc42-GTP
• ProvidingreliableresultswithconsistentreproducibilitythanotherCdc42activationassaykitsbasedoncertaindownstreamsignalingpathways
• Providingshorterexperimentaltime
• CouldbeusedtomonitortheactivationofCdc42incellsandintissuesbyimmunohistochemistry

ProductDescription

SmallGTPasesareasuper-familyofcellularsignalingregulators.Cdc42belongstotheRhosub-familyofGTPasesthatregulatecellmotility,celldivision,andgenetranscription.GTPbindingincreasestheactivityofCdc42,andthehydrolysisofGTPtoGDPrendersitinactive.

CurrentlytheactivationofCdc42proteinsisassayedwiththebindingofGTP-boundCdc42tothep21-bindingdomain(PBD)ofp21-activatedproteinkinase(PAK).Thismethodisbasedontheobservationthattheactive,GTP-boundCdc42(andRac)couldbindtothePBDofPAK.However,thereproducibilityofthismethodispoor.ThisispartiallyduetotherelativelyquickhydrolysisofGTPtoGDPduringtheassayprocedure,andthelowbindingaffinityofPBDtoCdc42-GTP.

NewEastBiosciencesCdc42ActivationAssayKitisbasedontheconfiguration-specificmonoclonalantibodythatspecificallyrecognizesCdc42-GTP,butnotCdc42-GDP.Giventhehighaffinityofmonoclonalantibodiestotheirantigens,theactivationassaycouldbeperformedinamuchshortertime.Thisassayprovidesthereliableresultswithconsistentreproducibility.

Theseanti-Cdc42-GTPmonoclonalantibodycanalsobeusedtomonitortheactivationofCdc42incellsandintissuesbyimmunohistochemistry.

NewEastBiosciencesCdc42ActivationAssayKitprovidesasimpleandfastmethodtomonitortheactivationofCdc42.Eachkitprovidessufficientquantitiestoperform20assays.

AssayPrinciple

NewEastBiosciencesCdc42ActivationAssayKitbasesontheconfiguration-specificanti-Cdc42-GTPmonoclonalantibodytomeasuretheactiveCdc42-GTPlevels,eitherfromcellextractsorfrominvitroGTPγSloADIngCdc42activationassays.Briefly,anti-activeCdc42mousemonoclonalantibodywillbeincubatedwithcelllysatescontainingCdc42-GTP.TheboundactiveCdc42willthenbepulleddownbyproteinA/Gagarose.TheprecipitatedactiveCdc42willbedetectedbyimmunoblotanalysisusinganti-Cdc42rabbitpolyclonalantibody.

   ProductManual
   ProtocolforIF/IHC

RelatedProducts
Catalog#	Name	Size	Price	DataSheet
80701	Cdc42ActivationAssayKit	20assays	$637
26905	ActiveCdc42-GTPMonoclonalAntibody	30μL	$495
26008	Anti-Cdc42MouseMonoclonalAntibody	100μL	$275
21010	Anti-Cdc42RabbitPolyclonalAntibody	100μL	$275

定制肽合成NewEast Biosciences为全球生物研究界提供高质量的粗制和纯化肽或拟肽。我们使用先进的自动，半自动和手动合成器以及Fmoc技术合成肽。肽酰胺键通过HOAT / HATU偶联形成。我们的肽通过制备型HPLC纯化，并且每个级分均通过使用不同溶剂系统的分析型HPLC进行进一步分析。收集并冻干仅通过质谱法验证的具有单个峰的级分。肽通常在1-2周内递送。粗肽也可以大量节省（参见价格表）。周转时间为5-7天。除了正常的肽合成外，我们还合成修饰的肽和多种抗原性肽（MAPs）。修饰在C末端，N末端和内部残基处进行。在C末端，氨基可与羧酸，异氰酸酯和磺酰氯反应形成酰胺，脲和磺酰胺。在此终端进行经典的生物素化，荧光标记和脂质化。这些修饰以及磷酸化和糖基化也可以在内部残基处进行。相反，在C末端的修饰非常具有挑战性，需要溶液阶段的干预。为了在C端进行修饰，必须将肽从树脂中释放出来。然后，用相应的化学方法修饰释放的肽。但是，我们已经开发了适当的化学方法，可以在固相的C末端引入生物素，荧光染料和胺。随着化学，我们降低了合成肽以及昂贵试剂的成本。作为NewEast的客户，我们将节省下来的钱转嫁给您。

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Publications:

1.  VibrioparahaemolyticusEffectorProteinsSuppressInflammasomeActivationbyInterferingwithHostAutophagySignaling    PLoSPathog.2013Jan;9(1):e1003142

2.  Interferongammainducesactinpolymerization,Rac1activationanddownregulatesphagocytosisinhumanmonocyticcells    Cytokine.2012Jan;57(1):158-68

3.  Invadopodiaandrolling-typemotilityarespecificfeaturesofhighlyinvasivep190bcr-ablleukemiccells    EuropeanJournalofCellBIOLOGyVolume91,Issues11–12,November–December2012,Pages978–987

4.  EpithelialjunctionformationrequiresconfinementofCdc42activitybyanovelSH3BP1complex    JCellBiol.2012Aug20;198(4):677-93

5.  PolarizedmigrationoflymphaticendothelialcellsiscriticallydependentonpodoplaninregulationofCdc42    AmJPhysiolLungCellMolPhysiol.2011Jan;300(1):L32-42

6.  TheAarskog-ScottSyndromeProteinFgd1RegulatesPodosomeFormationandExtracellularMatrixRemodelinginTransformingGrowthFactorbeta-StimulatedAorticEndothelialCells    MolCellBiol.2011Nov;31(22):4430-41

7.  MMP-9andCXCL8/IL-8ArePotentialTherapeuticTargetsinEpidermolysisBullosaSimplex    PLoSOne.2013Jul19;8(7):e70123

8.  p21-ActivatedKinase(PAK)RegulatesCytoskeletalReorganizationandDirectionalMigrationinHumanNeutrophils    PLoSOne.2013Sep3;8(9):e73063

9.  TheCdc42GuanineNucleotideExchangeFactorFGD6CoordinatesCellPolarityandEndosomalMembraneRecyclinginOsteoclasts    JBiolChem.2014Jun27;289(26):18347-59

10.  Cdc42InhibitorandUsesThereof    UnitedStatesPatentApplication20140194451

11.  Theα4NicotinicReceptorPromotesCD4+T-CellProliferationandaHelperT-CellImmuneResponse    MolecularPharmacologyJanuary2014vol.85no.150-61