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protein interactions - (Apr/19/2011 )

dear All,

My protein actb is a member of Nu4A complex. Using IPA, I found out that myc is interacting with some of the members of this complex but co ip studies did not show any interaction between myc and actb. Is it possible to pull down actb and myc along with other members of the complex? if yes how?



Hi, unfortunately I have no idea what IPA is...
BUT, maybe you could pull down your complex with anti-myc antibody? and then use proteinA (or G-this depends on the type of AB)-coupled sepharose beads to enrich the complex. The proteinA binds to the antibody-Fc portion. Slow centrifugation is used to separate the complex bound to antibody-sepahrose beads and the other stuff in the suspension.
Still it would be difficult to identify an unknown binding partner :/
good luck


IPA = immmunoprecipitation assay... yes it is possible if the conditions are correct and there is a complex of proteins formed to be pulling down one interaction and also get other proteins from the same complex.