ApplicationNotes | Immunoblotting:1:1,000-1:2,000ondenaturedprotein.Westernblot:1:1000-1:5000.Recognizesa220-240kDAbandondenaturingSDSgels.Thereisalsoa60-64kDabandpresentinmostlysates,theidentityofthissmallerfragmentisunknownbutitisnotseeninlysatesnotcontaininghumannestinprotein.
WI-38(ATCC#CCL75)canbeusedasapositivecontrol.U251celllineisalsopositive.
Immunocytochemistry:1:200oncellsfixedin4%paraformaldehydeandpermeABIlizedwith0.02%Triton
Immunohistochemistry:paraformaldehydeisthepreferredfixative:1:200onformalinfixed,paraffinembeddedtissuesections.Usetriton0.02%inblockonly;dilutionbuffer0.1%BSA;incubations1-2hoursRT.Humanfetalbrainissuggestedasapositivecontrol.(CAMessametal,Develop.BrainRes,134:87-92,2002;Gu,etal.,BrainRes.943:174-180,2002).Filamentousstaininginthecytoplasmofcells
Optimalworkingdilutionsmustbedeterminedbyenduser. |