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Heat-mediated antigen retrieval is recommended prior to staining, using a 10mM citrate buffer, pH 6.0, for 10 minutes followed by cooling at room temperature for 20 min. Following antigen retrieval, incubate samples with primary antibody for 10 min at room temperature. A suggested positive control is liver carcinoma.
TRADD (Tumor necrosis factor receptor type 1-associated DEATH domain protein) is an important mediator of FAS and TNFR1 associated apoptotic signaling. The nuclear form acts as a tumor suppressor by preventing ubiquitination and degradation of isoform p19ARF/ARF of CDKN2A by TRIP12. TRADD acts by interacting with TRIP12 and leading to the disruption of the interaction between TRIP12 and isoform p19ARF/ARF of CDKN2A. TRADD acts as an adapter molecule for TNFRSF1A/TNFR1 that specifically associates with the cytoplasmic domain of activated TNFRSF1A/TNFR1 mediating its interation with FADD. Overexpression of TRADD leads to two major TNF-induced responses - apoptosis and activation of NF-kappa-B.
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