ApplicationNotes | ELISAat1:400-1:3,200.
RIAat1:50-1:6,400.
Immunohistochemistry:Thisantibodyonlyreactswithdigestedmaterial.FixedtissuemustbedigestedwithchondroitinaseABC(Sigma). TheReference23757342shouldbeindicatedforIHC.(Reine,TM.,etal.,PubMedID:23757342)
WesternBlot:1:1000-1:4000usingECLdetectionmethod.ChondroitinaseABCtreatmentoflysatesampleishelpful. Inanyalkalibuffer,e.g.Trisoracetate(50mMor100mM)pH7.4-7.6.MakeastocksolutionbydilutingonevialofSigmaChondroitinaseA,B,C,(5units/vial)in100μL.Foruse,dilutestock10-foldintothebufferofchoice.Useataratioof50-100mgofproteoglycanper5unitsofenzyme.Onfixedcells,diluteinTrisacetateandonnon-fixedcellsydilute10-25foldinDMEM. Poole,C.A.,etal.,J.Histochem.Cytochem.1991.39:1175-1187.
Optimalworkingdilutionsmustbedeterminedbytheenduser. |