LyophilizedPowderLyophilizedPowder
Thisproductisfreezedried.Allwatermoleculeshavebeenremoved.
AntigenIncl.AntigenIncl.
ThisantibodyisshippedwithitsantigenFREEofcharge!
ShippedatRoomTemp.ShippedatRoomTemp.
Thislyophilizedproductisshippedatroomtemperature.Pleaseseeitscertificateofanalysisforfurtherstorageinstructions.
Type:Polyclonal
Source:Rabbit
Reactivity:h,m,r
ImmunogenPeptide(C)DYTWNKDNWTFSP,correspondingtoaminoacidresidues72-84ofhumanGPR65(AccessionQ8IYL9).1stextracellularloop.
Accession(Uniprot)NumberQ8IYL9
GeneID8477
PeptideconfirmationConfirmedbyaminoacidanalysisandmassspectrometry.
HomologyMouse–12/13aminoacidresiduesidentical;rat-11/13aminoacidresiduesidentical.
PurityAffinitypurifiedonimmobilizedantigen.
FormulationLyophilizedpowder.Reconstitutedantibodycontainsphosphatebufferedsaline(PBS),pH7.4,1%BSA,0.05%NaN3.
StoragebeforereconstitutionTheantibodyshipsasalyophilizedpowderatroomtemperature.Uponarrival,itshouldbestoredat-20°C.
Reconstitution25µl,50µlor0.2mldoubledistilledwater(DDW),dependingonthesamplesize.
Antibodyconcentrationafterreconstitution0.8mg/ml.
StorageafterreconstitutionThereconstitutedsolutioncanbestoredat4°Cforupto1week.Forlongerperiods,smallaliquotsshouldbestoredat-20°C.Avoidmultiplefreezingandthawing.Centrifugeallantibodypreparationsbeforeuse(10000xg5min).
ControlantigenstoragebeforereconstitutionLyophilizedpowdercanbestoredintactatroomtemperaturefor2weeks.Forlongerperiods,itshouldbestoredat-20°C.
Controlantigenreconstitution100µldoubledistilledwater(DDW).
Controlantigenstorageafterreconstitution-20°C.
PreadsorptionControl1µgpeptideper1µgantibody.
StandardqualitycontrolofeachlotWesternblotanalysis.
Applications:ifc,ih,wb
Mayalsoworkin:ic,ip,lci
WesternblotWesternblotanalysisofhumanJurkatT-cellleukemia(lanes1and4),humanTHP-1monocyticleukemia(lanes2and5)andhumanHL-60promyelocyticleukemia(lanes3and6)celllinelysates:
1-3. Anti-GPR65(TDAG8)(extracellular) Antibody(#AGR-043),(1:200).
4-6.Anti-GPR65(TDAG8)(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.
Westernblotanalysisofrat(lanes1and3)andmouse(lanes2and4) brainmembranes:
1,2. Anti-GPR65(TDAG8)(extracellular) Antibody(#AGR-043),(1:200).
3,4.Anti-GPR65(TDAG8)(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.
ImmunohistochemistryExpressionofGPR65inratfornix
Immunohistochemicalstainingofimmersion-fixed,freefloatingratfrozenbrainsectionsusing Anti-GPR65(TDAG8)(extracellular) Antibody(#AGR-043),(1:1200),followedbygoat-anti-rabbit-AlexaFluor-488.GPR65staining(green)appearsinglialprocesses(verticalarrows)andinthesideoffornix(FX)facingtheventricle(V,horizontalarrow).CellnucleiarestainedwithDAPI(blue).
IndirectflowcytometryIndirectflowcytometryofliveintacthumanJurkatT-cellleukemiacells:
___ Cells.
___ Cells+goat-anti-rabbit-FITC.
___ Cells+ Anti-GPR65(TDAG8)(extracellular) Antibody(#AGR-043),5µg+goat-anti-rabbit-FITC.
ReferencesJustus,C.R.etal.(2013)Front.Physiol.4,354.
Justus,C.R.etal.(2017)J.Transl.Med.15,204.
ScientificbackgroundGPR65,alsocalledTDAG8(T-celldeathassociatedgene8protein),isapH-sensingG-proteincoupledreceptor(GPCR)activatedbyacidicextracellularpHthroughtheprotonationofseveralhistidineresiduesinthereceptor’ssequence.GPR65isinvolvedincancercellmetastasisandproliferation,immunecellfunction,inflammation,andbloodvesselformation1.
HumanGPR65genehasbeenmappedtoalocationthatisassociatedwithTcelllymphomaandleukemiaabnormalities.
GPR65isprimarilyexpressedinimmunecellsandleukocyte-richtissuessuchascirculatingperipheralleukocytes,spleen,thymus,andtonsils.ExpressionisalsodetectedpainrelevantlocisuchasthedorsalrootganglianeuronsandparticularlysmalldiameterneuronsresponsIBLefornociception1,2.
GPR65hastheABIlitytoaffecttumordevelopmentandgrowth.OverexpressionofGPR65inLewislungcarcinomacellsincreasestumorgrowthinmiceandmayfacilitateresistancetoacidosis-mediatedcelldeath invitro throughproteinkinaseA(PKA)andERKrelatedpathways1.
KnockdownofGPR65inNCI-H460humannon-smallcelllungcancercellsdecreasescellsurvivalinacidiccondition.AnotherstudyshowsthatactivationofGPR65byacidosiscanpromoteevasionofcellapoptosisunderglutaminestarvationanditsoverexpressionhasbeenreportedtotransformimmortalizedmammaryepithelialcells1.
LyophilizedPowderControlAntigenIncluded
Clickheretoreceivea25µlfreetrialsample!
Lastupdate:11/12/2018
Anti-GPR65(TDAG8)(extracellular)Antibody(#AGR-043)isahighlyspecificantibodydirectedagainstanepitopeofthehumanprotein.Theantibodycanbeusedinwesternblot,immunohistochemistry,andlivecellflowcytometry.IthasbeendesignedtorecognizeGPR65fromrat,mouse,andhumansamples.
Forresearchpurposesonly,notforhumanuse
WesternblotWesternblotanalysisofhumanJurkatT-cellleukemia(lanes1and4),humanTHP-1monocyticleukemia(lanes2and5)andhumanHL-60promyelocyticleukemia(lanes3and6)celllinelysates:
1-3. Anti-GPR65(TDAG8)(extracellular) Antibody(#AGR-043),(1:200).
4-6.Anti-GPR65(TDAG8)(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.
Westernblotanalysisofrat(lanes1and3)andmouse(lanes2and4) brainmembranes:
1,2. Anti-GPR65(TDAG8)(extracellular) Antibody(#AGR-043),(1:200).
3,4.Anti-GPR65(TDAG8)(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.
