A practical guide to fractionating cells, validating nuclear and cytoplasmic purity, normalizing p65 signals, troubleshooting contamination, and limiting conclusions to what an NF-κB translocation assay can establish.
Constitutive NF-κB activation is an experimental conclusion, not a synonym for one positive assay. This guide explains the evidence needed to demonstrate persistent baseline signaling and pathway dependence in cancer models.
A mechanism-first explanation of how TNF-α activates canonical NF-κB through TNFR1, ubiquitin-organized signaling, TAK1, IKK, and IκBα degradation.
A mechanism-focused guide to the canonical NF-κB p50/RelA heterodimer, from subunit composition and DNA binding to localization and assay interpretation.