CrossreactivityofthemonoclonalantibodydirectedagainstH3K27ac
TotestthespecificityanELISAwasperformedusingaserialdilutionofthemonoclonalantibodyagainstH3K27ac(Cat.No.25238).ThewellswerecoatedwithpeptidescontainingtheunmodifiedH3K27regionaswellastheacetylatedH3K27andtheacetylatedH3K9.Figure1showsahighspecificityoftheantibodyforthepeptidecontainingthemodificationofinterest.
ImmunofluorescenceusingthemonoclonalantibodydirectedagainstH3K27ac
HeLacellswerestainedwiththeantibodyagainstH3K27ac(Cat.No.25238)andwithDAPI.Cellswerefixedwith4%formaldehydefor10minutesandblockedwithPBS/TX-100containing5%normalgoatserumand1%BSA.ThecellswereimmunofluorescentlylabelledwiththeH3K27acantibody(left)diluted1:500inblockingsolutionfollowedbyananti-mouseantibodyconjugatedtoAlexa594.ThemiddlepanelshowsstainingofthenucleiwithDAPI.Amergeofthetwostainingsisshownontheright.
Storeat–80°Cforupto2years.Centrifugeafterfirstthawtomaximizeproductrecovery.Aliquottoavoidrepeatedfreeze/thawcycles.Aliquotsmaybestoredat–20°Cforatleastonemonth.
ELISA(1:3000)
WB(1:1000–1:2000)
IF(1:500)