| ApplicationNotes | ImmunohistochemistryAnalysis:A1:500dilutionfromarepresentativelotdetectedGABAinratbraintissue. 
 ELISAAnalysis:ArepresentativelotfromanindependentlaboratorydetectedGABAinacompetitiveELISA.
 
 ImmunohistochemistryProtocol:
 
 Tissuesfixedwith4%paraformaldehydeand0-0.5%glutaraldehydegivesgoodresults.Glutaraldehydeisrequiredforantibodyreactivity.
 
 1)Tissueisfixedwith4%paraformaldehyde,0-0.5%glutaraldehyde,0.5%potassiumdichromatein0.1MphosphatebufferatpH6.5.
 
 2)Tissueispost-fixedovernight,vibratomesectionedin50mmandincubatedin0.05MTrisbuffer,pH6.5forthreehours.
 
 3)Sectionsareincubatedfor18-24hoursinAB175dilutedinPBScontaining0.1%sodiumazide,0.2%TritonX-100and1%normalgoatserum.
 
 4)FluoresceinconjugatedantibodyorABCsystemmaybeusedasthesecondaryreagent.
 
 Note:Withoutcolchicinepretreatmentwell-stainedcellbodiesarevisIBLeinthecerebralcortex,cerebrallarcortex,superiorcolliculusandsomebrainstemraphe.Withcolchicinepretreatment,additionalcellbodystainingispresentintheinterpeduncularnucleusandthedorsalcolumnnuclei.
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