My experiment uses immunocytochemistry to observe the translocation of NFkB from the cytoplasm into the nucleus. In most articles, the primary antibody is NFkB p65. I am wondering if anyone used NFkB p50 to check translocation of NFkB after TNF alpha treatment? In my opinion, p65 and p50 subunits are translocated into nucleus each other.
Dear Tran Hue Vy An,
p50 of NF-kB can associate with different partners and it can form homodimers, which can also translocate.However, p50 does not have a transactivation domain and cannot on its own activate gene expression- rather in the form of homodimers it can block.
We have looked at translocation of p50 in WB and have published on translocation of the related p52 into the nucleus (Biol Chemistry, 380: 1193, 1999). However, this translocation does not tell you whether or not you induce gene expression, since this is dependent on the active principle which is p65. p65 is the active priniciple because it has a transactivation domain (TAD). Therefore most people prefer to study transloaction of p65 when they stimulate cells.
Yours
Loems
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