Forinductionofc-fosproteinactivityratswereinjectedwith1.0mlof1.5MNaClper100gramsofbodyweight.Negativecontrolratswereinjectedwiththesamevolumeofnormalsaline.TheImmunoStarc-fosantiserumwasqualitycontroltestedusingstandardimmunohistochemicalmethods.Theantiserumdemonstratesstronglypositivelabelingofratparaventricularnucleusandsupraopticnucleususingindirectimmunofluorescentandbiotin/avidin-HRPtechniques.Nolabelingwasseeninnegativecontrolrats.Recommendedprimarydilutionsforthesemethodsare1/200-1/400inPBS/0.3%TritonX-100-FITCTechniqueand1/4000-1/6000inPBS/0.3%TritonX-100-biotin/avidin-HRPTechnique.

InjectionofintraperitonealhypertonicsalinetoprovokefosexpressionismostusuallyusedtosetICCstainingparameters. Itiscriticalthattissueisharvested60-90minutespostinjectionforthisandanyotherstressthatisused. Inject1ml/100gbodyweight1.5MNaClintraperitoneally. Harvestbrains90minutespostinjection. Timeiscritical.

Specificityoftheantiserumwasdemonstratedbyblockageofstaininginexperimentalratsbyomissionofc-fosantibodyorbysubstitutionofantibodypre-incubatedwithsyntheticpeptideortheconjugate.Immunoblotanalysisofmediobasalhypothalamusshowedasinglebandofapproximately55-60kD.

PhotoDescription: IHCimageofneuronsstainingforc-Fosintheratsupraopticnucleus.Thetissuewasfixedwith4%formaldehydein0.1Mphosphatebuffer,beforebeingremovedandpreparedforvibratomesectioning.FloatingsectionswereincubatedatRTin10%goatseruminPBS,beforestandardIHCprocedure.Primaryantibodywasincubatedat1:5000for48hours,goatanti-rabbitsecondarywassubsequentlyaddedfor1hourafterwashingwithPBS.Lightmicroscopystainingwasachievedwithstandardbiotin-streptavidin/HRPprocedureandDABchromogen.Thesectionwasthenmountedonslideswithpermount.

Host:Rabbit

Quantity/Volume:100µL

State:LyophilizedWholeSerum

ReactsWith:Mouse,Rat

AvailABIlity:InStock

AlternateNames: Proto-oncogenec-Fos;Cellularoncogene;G0/G1switchregulatoryprotein7;Proto-oncogeneprotein;G0S7;p55;AP-1;FBJmurineosteosarcomaviraloncogenehomolog,anti-CFOS

GeneSymbol/ID,Accession#: FOS,2353

RRID: AB_572267