LyophilizedPowderLyophilizedPowder
Thisproductisfreezedried.Allwatermoleculeshavebeenremoved.
AntigenIncl.AntigenIncl.
ThisantibodyisshippedwithitsantigenFREEofcharge!
ShippedatRoomTemp.ShippedatRoomTemp.
Thislyophilizedproductisshippedatroomtemperature.Pleaseseeitscertificateofanalysisforfurtherstorageinstructions.
Type:Polyclonal
Source:Rabbit
Reactivity:h,m,r
ImmunogenPeptide(C)DYTWNKDNWTFSP,correspondingtoaminoacidresidues72-84ofhumanGPR65(AccessionQ8IYL9).Extracellular,1stloop.
Accession(Uniprot)NumberQ8IYL9
GeneID8477
PeptideconfirmationConfirmedbyaminoacidanalysisandmassspectrometry.
HomologyMouse–12/13aminoacidresiduesidentical;rat-11/13aminoacidresiduesidentical.
PurityAffinitypurifiedonimmobilizedantigen.
FormulationLyophilizedpowder.Reconstitutedantibodycontainsphosphatebufferedsaline(PBS),pH7.4,1%BSA,0.05%NaN3.
LabelFluoresceinisothiocyanate(FITC).
StoragebeforereconstitutionTheantibodyshipsasalyophilizedpowderatroomtemperature.Uponarrival,itshouldbestoredat-20°C.
Reconstitution50µldoubledistilledwater(DDW).
Antibodyconcentrationafterreconstitution1mg/ml.
StorageafterreconstitutionThereconstitutedsolutioncanbestoredat4°C,protectedfromthelight,forupto1week.Forlongerperiods,smallaliquotsshouldbestoredat-20°C.Avoidmultiplefreezingandthawing.Centrifugeallantibodypreparationsbeforeuse(10000×g5min).
ControlantigenstoragebeforereconstitutionLyophilizedpowdercanbestoredintactatroomtemperaturefor2weeks.Forlongerperiods,itshouldbestoredat-20°C.
Controlantigenreconstitution100µldoubledistilledwater(DDW).
Controlantigenstorageafterreconstitution-20°C.
PreadsorptionControl1µgpeptideper1µgantibody.
StandardqualitycontrolofeachlotWesternblotanalysis(unlabeledantibody,#AGR-043),anddirectflowcytometry(labeledantibody).
Applications:fc
Mayalsoworkin:ic,lci
DirectflowcytometryDirectflowcytometryofliveintacthumanJurkatT-cellleukemiacells:
___ Cells.
___ Cells+rabbitIgGisotypecontrol-FITC.
___ Cells+Anti-GPR65(TDAG8)(extracellular)-FITCAntibody (#AGR-043-F),(5µg).
DirectflowcytometryofliveintactmouseM1myeloidleukemiacells:
___ Cells.
___ Cells+rabbitIgGisotypecontrol-FITC.
___ Cells+Anti-GPR65(TDAG8)(extracellular)-FITCAntibody(#AGR-043-F),(5µg).
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.Expressionisalsodetected pain relevantlocisuchasthedorsalrootganglianeuronsandparticularlysmalldiameterneuronsresponsIBLefornociception1,2.
GPR65hastheABIlitytoaffecttumordevelopmentandgrowth.OverexpressionofGPR65inLewislungcarcinomacellsincreasestumorgrowthinmiceandmayfacilitateresistancetoacidosis-mediatedcelldeath invitro throughproteinkinaseA(PKA)andERKrelatedpathways1.
KnockdownofGPR65inNCI-H460humannon-smallcelllungcancercellsdecreasescellsurvivalinacidiccondition.AnotherstudyshowsthatactivationofGPR65byacidosiscanpromoteevasionofcellapoptosisunderglutaminestarvationanditsoverexpressionhasbeenreportedtotransformimmortalizedmammaryepithelialcells1.
LyophilizedPowderControlAntigenIncluded
Lastupdate:20/03/2019
Anti-GPR65(TDAG8)(extracellular)Antibody(#AGR-043)isahighlyspecificantibodydirectedagainstanepitopeofthehumanprotein.Theantibodycanbeusedinwesternblot, immunohistochemistry,andlivecellflowcytometry.IthasbeendesignedtorecognizeGPR65fromrat,mouse,andhumansamples.
Anti-GPR65(TDAG8)(extracellular)-FITCAntibody(#AGR-043-F)isdirectlyconjugatedtofluoresceinisothiocyanate(FITC).Thislabeledantibodycanbeusedinimmunofluorescentapplicationssuchasdirectlivecellflowcytometry.
Forresearchpurposesonly,notforhumanuse
DirectflowcytometryDirectflowcytometryofliveintacthumanJurkatT-cellleukemiacells:
___ Cells.
___ Cells+rabbitIgGisotypecontrol-FITC.
___ Cells+Anti-GPR65(TDAG8)(extracellular)-FITCAntibody (#AGR-043-F),(5µg).
DirectflowcytometryofliveintactmouseM1myeloidleukemiacells:
___ Cells.
___ Cells+rabbitIgGisotypecontrol-FITC.
___ Cells+Anti-GPR65(TDAG8)(extracellular)-FITCAntibody(#AGR-043-F),(5µg).
ReferencesJustus,C.R.etal.(2013)Front.Physiol.4,354.
Justus,C.R.etal.(2017)J.Transl.Med.15,204.