货号:80301
CAS号:80301
供应商:NewEastBio
数量:30000
英文名:GaiActivationAssayKit
保质期:两年
保存条件:-20℃
注册证号:80301
规格:20T
GαiActivationAssayKit[80301]
[20assays]
 

ProductHighlight

  • Configuration-specificmonoclonalantibodythatrecognizesGαi-GTP,butnotGαi-GDP
  • TheonlyavailableGαi activationassaykitdirectlymeasuresthelevelofGαi-GTP
  • ProvidingreliableresultswithconsistentreproducibilitythanotherGαi activationassaykitsbasedoncertaindownstreamsignalingpathways
  • Providingshorterexperimentaltime
  • CouldbeusedtomonitortheactivationofGαi incellsandintissuesbyimmunohistochemistry

ProductDescription

Astructurallydiverserepertoireofligands,fromphotonstolargepeptides,activatesGprotein-coupledreceptors(GPCRs)toelicittheirphysiologicalfunctions.Ligand-boundGPCRs,inturn,functionasguaninenucleotideexchangefactorscatalyzingtheexchangeofGDPboundontheGαsubunitwithGTPinthepresenceofGβγ,causingthedissociationoftheGαsubunitfromtheGβγdimertoformtwofunctionalunits(GαandGβγ).BothGαandGβγsubunitssignaltovariouscellularsignalingpathways.Basedonthesequenceandfunctionalhomologies,Gproteinsaregroupedintofourfamilies:Gs,Gi,Gq,andG12.

i familyisthelargestfamilyofGproteins.TheyrelaysignalsfrommanyGPCRstoregualtevariousBIOLOGicalfunctions.TherewerenodirectmethodstomeasuretheactivationofGαi proteinsbyreceptors(untilthisassaykit).Mostreportsusedoneofthedownstreampathway,i.e.theinhibitionofadenylylcyclases,asareadout.Alternatively,sensitivitytopertussistoxin(PTX)wasusedasanindicatorofpossIBLeGαi proteinsinvovledinasignalingpathway.

NewEastBiosciencesGαi ActivationAssayKitprovidesadirectmeasurementoftheactivationofGαi proteins.ThisisasimpleandfasttooltomonitortheactivationofGαi.Eachkitprovidessufficientquantitiestoperform20assays.

NewEastBiosciencesGαi ActivationAssayKitisbasedonthemonoclonalantibodyspecificallyrecognizingtheactiveGTP-boundGαi proteins.ThismonoclonalantibodyhasmuchloweraffinitytowardstheinactiveGαi proteins.Therefore,afteractivationbyreceptorsignals,activeGTP-boundGαi proteinscouldbeimmunoprecipitatedbythismonoclonalantibodyandfurtherquantifiedbywesternblotwithanotheranti-Gαi antibody.

 

    AssayPrinciple

    NewEastBiosciencesGαi ActivationAssayKitisanimmunoprecipitation/westernblotassaytomeasurethelevelsofactiveGTP-boundGαi proteins,eitherfromcellextractsorfrominvitroGTPγSloadedGαiproteins.Briefly,theanti-activeGαi monoclonalantibodywillspecificallybindtoactiveGαi protein.Thisantibody/Gαi complexwillthenbepulleddownbyproteinA/Gagarose.TheprecipitatedactiveGαi proteinswillbedetectedbyimmunoblotswithanotheranti-Gαi antibody.

       

      Publications:
      1.  IdentificationofnovelsignallingrolesandtargetsforGalphaandGbetagammadownstreamoftheinsulin-likegrowthfactor1receptorinvascularsmoothmusclecells
          BiochemJ.2013Feb15;450(1):209-19
      2.  CRHactivationofdifferentsignalingpathwaysresultsindifferentialcalciumsignalinginhumanpregnantmyometriumbeforeandduringlabor
          JClinEndocrinolMetab.2012Oct;97(10):E1851-61
      3.  CXCR4activationdefinesanewsubgroupofSonichedgehog-drivenmedulloblastoma
          CancerRes.2012Jan1;72(1):122-32
      4.  RegulationofestrADIolandprogesteroneproductionbyCRH-R1and-R2isthroughdivergentsignalingpathwaysinculturedhumanplacentaltrophoblasts
          Endocrinology.2012Oct;153(10):4918-28
      5.  WNT-5AstimulatestheGDP/GTPexchangeatpertussistoxin-sensitiveheterotrimericGproteins
          CellSignal.2011Mar;23(3):550-4
      6.  HeterotrimericGalpha(i)proteinsareregulatedbylipopolysaccharideandareanti-inflammatoryinendotoxemiaandpolymicrobialsepsis
          BiochimBiophysActa.2011Mar;1813(3):466-72
      7.  Estrogen-andxenoestrogen-inducedERKsignalinginpituitarytumorcellsinvolvesestrogenreceptor-αinteractionswithGprotein-αiandcaveolinI
          SteroidsVolume77,Issue5,April2012,Pages424–432
      8.  Na/HexchangerregulatoryfactorscontrolparathyroidhormonereceptorsignalingbyfacilitatingdifferentialactivationofG(alpha)proteinsubunits
          JBiolChem.2010Aug27;285(35):26976-86
      9.  Beta-arrestin-butnotGprotein-mediatedsignalingbythe"decoy"receptorCXCR7
          ProcNatlAcadSciUSA.2010Jan12;107(2):628-32
      10.  StructuralbasisforactivationoftrimericGiproteinsbymultiplegrowthfactorreceptorsviaGIV/Girdin
          MolBiolCell.2014Nov5;25(22):3654-71
      11.  CanonicalandNon-canonicalG-proteinSignalingHelpsCoordinateActinDynamicstoPromoteMacrophagePhagocytosisofZymosan
          MolCellBiol.2014Nov15;34(22):4186-99
      12.  FattyAcid-bindingProtein5(FABP5)RegulatesCognitiveFunctionBothbyDecreasingAnandamideLevelsandbyActivatingtheNuclearReceptorPeroxisomeProliferator-activatedReceptorβ/δ(PPARβ/δ)intheBrain
      JBiolChem.2014May2;289(18):12748-58
       
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