Deprecated: Required parameter $cat_id follows optional parameter $type in /www/wwwroot/ebimall.com/systems/hong.php on line 2088

Deprecated: Required parameter $where follows optional parameter $tree_id in /www/wwwroot/ebimall.com/systems/hlb.php on line 3505
Antibodies-Online/GST-Trap M/ABIN1082192/200 tests188bio精品生物—专注于实验室精品爆款的电商平台 - 蚂蚁淘旗下精选188款生物医学科研用品
您好,欢迎您进入188进口试剂采购网网站! 服务热线:4000-520-616
蚂蚁淘商城 | 现货促销 | 科研狗 | 生物在线
产品资料

Antibodies-Online/GST-Trap M/ABIN1082192/200 tests

Antigen
GlutathioneSTransferase(GST)
  • GST
  • GST13-13
  • GST13
  • GST13-13
  • GSTK1-1
  • hGSTK1
  • GSTS
  • GSTS1-1
  • PGD2
  • PGDS
  • H-PGDS
  • Ptgds2
  • 1500002K10Rik
  • Gst
  • glutathioneS-transferasekappa1
  • hematopoieticprostaglandinDsynthase
  • microsomalglutathioneS-transferase1
  • GSTK1
  • HPGDS
  • Hpgds
  • Mgst1
Reactivity
Schistosomajaponicum
3Human
3Schistosomajaponicum
2Mammalian
2Mouse(Murine)
2Rabbit
2Rat(Rattus)
2Tag
1AllSpecies
1Chicken
1Cow(Bovine)
1Dog(Canine)
1Fish
1Goat
1GuineaPig
1Monkey
1Pig(Porcine)
1Sheep(Ovine)
Host
Camelidae
AntibodyType
RecombinantAntibody
Conjugate
MagneticParticles
Application
AffinityMeasurement(AM),ChromatinImmunoprecipitation(ChIP),EnzymeActivityAssay(EAA),Immunoprecipitation(IP),MassSpectrometry(MS),ProteinComplexImmunoprecipitation(Co-IP),Pull-DownAssay(Pull-Down),Purification(Purif)
Options
Bulkdiscount
Supplier
SupplierProductNo.
PurposeGST-TrapisahighlyspecificandrobustGST-bindingproteincoupledtoamonovalentmatrix(magneticparticles)forbiochemicalanalysisofGSTfusionproteinsandtheirinteractingpartners.
SampleTypeCellExtracts
SpecificityBindingcapacity:10µLGST-Trap_Mslurrybinds0.25-0.5µgofGST
CharacteristicsAntibodies-extremelypowerfultoolsinbiomedicalresearch-arelargecomplexmolecules(~150kDa)consistingoftwoheavyandtwolightchains.Duetotheircomplexstructure,theuseofantibodiesisoftenlimitedandhinderedbybatch-to-batchvariations.

Camelidae(camels,dromedaries,llamasandalpacas)possessfunctionalantibodiesdevoidoflightchains,so-calledheavychainantibodies(hcAbs).hcAbsrecognizeandbindtheirantigensviaasinglevariabledomain(VHH).TheseVHHdomainsarethesmallestintactantigenbindingfragments(~13kDa).

Nano-Trapsarebasedonsingledomainantibodyfragments(VHHs)derivedfromalpaca.
ComponentsGST-Trapcoupledtomagneticparticles
MaterialnotincludedLysisbuffer(CoIP),10xRIPAbuffer,Dilutionbuffer,Washbuffer,Elutionbuffer
AlternativeNameGST(GSTELISAKitAbstract)
BackgroundFusionofproteinstoGlutathioneS-Transferase(GST)ofSchistosomajaponicumisacommontechniquetoincreasesolubilityandexpressionlevelofaprotein.FurThermore,GST-taggedfusionproteinsareoftenusedinprotein-proteininteractionstudies.BothapproachesrequirehighlyspecifictoolstoisolateanddetectGSTfusionproteins
ResearchAreaTags/Labels,Metabolism,Immunology
ApplicationNotesForbiochemicalanalysesincludingmassspectroscopyandenzymeactivitymeasurementstheseGST-fusionproteinsandtheirinteractingfactorscanbeisolatedfastandefficiently(onestep)viaimmunoprecipitationusingtheGST-Trap.SincetheinteractionismediatedbyasmallGST-bindingproteincoupledtomagneticparticlestheGST-Trap_MenablespurificationofanyproteinofinterestfusedtoGST.
Comment

Beadsize0.5-1µm

AssayTime1.5h
Protocol
  • RobustandversatiletoolforbiochemicalanalysesofGST-fusionproteins
  • Shortincubationtimes(5-30min)
  • Lowunspecificbinding
  • Quantitativeisolationoffusionproteinsandtransientlyboundfactorsfromcellextractsororganelles
  • Nocontaminatingheavyandlightchainsofconventionalantibodies
ReagentPreparation

Suggestedbuffercomposition

  • Lysisbuffer:10mMTris/ClpH7.5,150mMNaCl
  • Dilutionbuffer:10mMTris/ClpH7.5,150mMNaCl
  • Washbuffer:10mMTris/ClpH7.5,150mMNaCl
  • Elutionbuffer:200mMglycinepH2.5

AssayProcedure
  • 1.ForoneimmunoprecipitationreactionresUSPendcellpelletof1mLbacteriacultureexpressingGSTin100-200µLlysisbufferbypipetting.
    optional:add1mMPMSFand0.1mg/mllysozymetolysisbuffer
  • 2.Rotatetubefor1hat4°C.
  • 3.Sonifycellpellettodisruptbacterialmembrane.
  • 4.Spincelllysateat20.000xgfor5-10minutesat4°C.
  • 5.Transfersupernatanttoapre-cooledtube.Adjustvolumewithdilutionbufferto500µL.Discardpellet.Forimmunoblotanalysisdilute30µLcelllysatewith10µL4xSDS-samplebuffer(->refertoasinput).
  • 6.EquilibrateGST-Trap_Mbeadsindilutionbuffer.Resuspendmagneticparticlesbyvortexingandtransfercalculatedvolume(20-30µL)inanewreactioncupwith250µLicecolddilutionbuffer.Magneticallyseparateuntilsupernatantisclearandwashtwicewith250µLofcolddilutionbuffer.
  • 7.AddcelllysatetoequilibratedGST-Trap_MbeadsandincubatetheGST-Trap_Mbeadswiththecelllysateunderconstantmixingfor10min–2hatroomtemperatureor4°C.
    note:duringincubationofproteinsamplewiththeGST-Trap_Mthefinalconcentrationofdetergentsshouldnotexceed0.2%toavoidunspecificbindingtothematrix
  • 8.Magneticallyseparateuntilsupernatantisclear.Forwesternblotanalysisdilute30µLsupernatantwith10µL2xSDS-samplebuffer(->refertoasnon-bound).Discardremainingsupernatant
  • 9.Washbeadstwotimeswith500µLicecoldwashbuffer.
    optional:increasesaltconcentrationinthesecondwashingstepupto500mM
  • 10.ResuspendGST-Trap_Mbeadsin100µL2xSDS-Samplebufferorgotostep11.
  • 11.Boilresuspendedbeadsfor10minutesat95°Ctodissociatetheimmunocomplexesfromthebeads.Thebeadscanbecollectedbycentrifugationat2.500xgfor2minutesat4°CandSDS-PAGEisperformedwiththesupernatant(->refertoasbound).
  • 12.optional:eluteboundproteinsbyadding50µL0.2MglycinepH2.5(incubationtime:30secunderconstantmixing)followedbycentrifugation.Transferthesupernatanttoafreshcupandadd5µL1MTrisbase(pH10.4)forneutralization.Toincreaseelutionefficiencythisstepcanberepeated.
RestrictionsForResearchUseonly
Concentration2.5mLresin
Buffer1xPBS,0.01%Sodiumazide
PreservativeSodiumazide
PrecautionofUseThisproductcontainssodiumazide:aPOISONOUSANDHAZARDOUSSUBSTANCEwhichshouldbehandledbytrainedstaffonly.
HandlingAdviceDonotfreeze.
Storage4°C
ExpiryDate12months
SupplierImages
Western Blotting (WB) image for GST-Trap M (ABIN1082190)PulldownofGSTusingtheGST-TrapImmunoprecipitations(IP)ofGSTfromproteinextr...
Western Blotting (WB) image for GST-Trap M (ABIN1082190)WesternBlotAnalysisofpurifiedGST:10,25,50,100,250,50ngGST,detectedwith...
Western Blotting (WB) image for GST-Trap M (ABIN1082190)ProteinlysatepfE.colicellsexpressingGSTwasspearatedbySDS-Pageandanalyzed...