| 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. |