Daxx Overexpression Lysate

HEK293T cells in 10-cm dishes were transiently transfected with a non-lipid polymer transfection reagent specially designed and manufactured for large volume DNA transfection. Transfected cells were cultured for 48hrs before collection. The cells were lysed in modified RIPA buffer (25mM Tris-HCl pH7.6, 150mM NaCl, 1% NP-40, 1mM EDTA, 1xProteinase inhibitor cocktail mix, 1mM PMSF and 1mM Na3VO4, and then centrifuged to clarify the lysate. Protein concentration was measured by BCA protein assay kit.

Daxx (death-domain-associated protein) was originally identified as a cytoplasmic protein that binds to the death domain of the transmembrane death receptor Fas. It is now that known that the a large porportion of Daxx molecules are nuclear and associate with the promyelocytic leukaemia nuclear body (PML-NB) and other subnuclear domains (reviewed in Salomoni and Khelifi). The promyelocytic leukemia protein Pml is an essential component of the PML-NB. Data suggests that certain stimuli including Fas stimulation, causes Daxx to be translocated from the nucleus to the cytoplasm. Daxx is ubiquitously expressed, and particularly high levels of Daxx have been reported in the thymus and testes. At the cellular level Daxx may be found in cytoplasm and within heterochromatic regions of the nucleus. Daxx has been reported to interact and co-localize with Pml in the nucleus of tumor cell lines and primary cells. Pml is necessary for Fas-induced cell death and Daxx pro-apoptotic functions. Daxx is thought to have both pro- and anti-apoptotic functions depending on the stimulus and the cell type. Daxx pro-apoptotic functions appear to occur in both the cytoplasm and nucleus. Although the mechanisms remain to be fully elucidated, research indicates that Daxx plays a role in the pathology of human diseases including cancer and neurodegerative disorders. Human Daxx is a 740 amino acid protein (GenBank no. gi/62898369).

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Lysates are guaranteed for 6 months from date of receipt.

Array NBP2-09623

• Daxx Antibodies
• Daxx Lysates
• Daxx Proteins