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Alomone/Anti-GPR4 (extracellular) Antibody/AGR-041/0.2 ml

Cat#:AGR-041
  • LyophilizedPowder
  • AntigenIncl.
  • ShippedatRoomTemp.
  • Type:Polyclonal
    Source:Rabbit
    Reactivity:h,m,r
    Immunogen
    Peptide(C)RDRYNHTF(S)FEKFPME,correspondingtoaminoacidresidues162-177ofratGPR4(Accession Q4KLH9).2nd extracellularloop.
    Accession(Uniprot)NumberQ4KLH9
    GeneID308408
    PeptideconfirmationConfirmedbyaminoacidanalysisandmassspectrometry.
    HomologyRat,mouse,human-15/16aminoacidresiduesidentical.
    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,wb
    Mayalsoworkin:ic,ih,ip,lci
    Westernblot
    Westernblotanalysisofratdorsalrootganglionlysate:
    1. Anti-GPR4(extracellular)Antibody(#AGR-041),(1:200).
    2.Anti-GPR4(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.
    Westernblotanalysisofrat(lanes1and3)andmouse(lanes2and4) kidneymembranes:
    1,2. Anti-GPR4(extracellular)Antibody(#AGR-041),(1:200).
    3,4.Anti-GPR4(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.
    Indirectflowcytometry
    IndirectflowcytometryofliveintacthumanTHP-1monocyticleukemiacells:
    ___Cells.
    ___Cells+goat-anti-rabbit-FITC.
    ___Cells+ Anti-GPR4(extracellular)Antibody (#AGR-041),(2.5µg)+goat-anti-rabbit-FITC.
    References
    1. Damaghi,M.etal.(2013)Front.Physiol. 4,370.
    2. Miyano,K.etal.(2014)J.Pharmacol.Sci. 126,302.
    3. Justus,C.R.etal.(2013)Front.Physiol.4,354.
    4. Im,D.S.(2005)ActaPharmacol.Sin.26,1435.
    Scientificbackground

    G-proteincoupledreceptor4(GPR4)belongstoaproteinfamilycomprisedof3relatedG-proteincoupledreceptors(GPCRs):GPR4,OGR1/GPR68andTDAG8/GPR65. LikeallGPCRs,GPR4"sstructureiscomposedofseven-transmembrane-spanningdomainslinkedbyanN-terminalextracellulardomainandaC-terminalintracellulardomain2.

    GPR4respondstopHchangesandisactivatedbyaphysiologicalincreaseinextracellularH+ throughseveralsignalingpathwaysincluding-theG12/13-protein/Rho,theGq/PLC,andtheGs-protein/cAMPpathway1,2.ItisfullyactivatedatpHlevelsof6.8andlowerandisinactiveatpHlevelsof7.5andhigher3,4.

    RecentstudiesshowthatthepH-sensingGPCRsalsohavetheABIlitytoregulatecancercellmetastasisandproliferation,immunecellfunction,inflammation,bloodvesselformation,endothelialbarrierfunctionthusmakingthemanimportanttherapeutictargetfordifferentdiseases. OverexpressionofGPR4inhumancancercellsshowsinvolvementindrivingormaintainingtumorformation.

    GPR4isexpressedinseveraltissuesincludinglung,vascularendothelial cells,kidney,heart,liver,skeletalmuscle,ovary,andplacenta.

    Applicationkey:

    CBE-Cell-basedELISA,FC-Flowcytometry,ICC-Immunocytochemistry,IE-IndirectELISA,IFC-Indirectflowcytometry,IHC-Immunohistochemistry,IP-Immunoprecipitation,LCI-Livecellimaging,N-Neutralization,WB-Westernblot

    Speciesreactivitykey:

    H-Human,M-Mouse,R-Rat
    receivea25µlfreetrialsample!
    Lastupdate:14/03/2019

    Anti-GPR4(extracellular)Antibody(#AGR-041)isahighlyspecificantibodydirectedagainstanepitopeoftheratprotein.Theantibodycanbeusedinwesternblotandindirectlivecellflowcytometryapplications.Theantibodyrecognizesanextracellularepitopeandisthusidealfordetectingthereceptorinlivingcells.IthasbeendesignedtorecognizeGPR4frommouse,rat,andhumansamples.

    Forresearchpurposesonly,notforhumanuse
    Westernblot
    Westernblotanalysisofratdorsalrootganglionlysate:
    1. Anti-GPR4(extracellular)Antibody(#AGR-041),(1:200).
    2.Anti-GPR4(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.
    Westernblotanalysisofrat(lanes1and3)andmouse(lanes2and4) kidneymembranes:
    1,2. Anti-GPR4(extracellular)Antibody(#AGR-041),(1:200).
    3,4.Anti-GPR4(extracellular)Antibody,preincubatedwiththecontrolpeptideantigen.
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