|Detection of Mouse Autoimmune Regulator/AIRE by Western Blot. Western blot shows lysates of mouse splenocytes and mouse thymus tissue. PVDF Membrane was probed with 1 µg/mL of Mouse Autoimmune Regulator/AIRE Monoclonal Antibody (Catalog # MAB6184) followed by HRP-conjugated Anti-Rat IgG Secondary Antibody (Catalog # HAF005). Specific bands were detected for Autoimmune Regulator/AIRE at approximately 70 and 48 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.|
Intracellular Staining by Flow Cytometry
Detection of Autoimmune Regulator/AIRE in Mouse splenocytes by Flow Cytometry. |
Mouse splenocytes were stained with Rat Anti-Mouse Autoimmune Regulator/AIRE Monoclonal Antibody (Catalog # MAB6184, filled histogram) or isotype control antibody (Catalog # MAB006, open histogram), followed by Phycoerythrin-conjugated Anti-Rat IgG Secondary Antibody (Catalog # F0105B). To facilitate intracellular staining, cells were fixed with paraformaldehyde and permeabilized with saponin.
AutoImmune REgulator (AIRE) is an approximately 60 kDa nuclear and cytosolic protein that is required for the development of T cell tolerance. It regulates the expression of self-antigens by thymic epithelial cells, and mutations in AIRE are causative of the autoimmune disorder, APECED. AIRE regulates gene transcription through interactions with DNA, histone H3, and the nuclear matrix. It contains one HSD domain (aa 1-105), a nuclear localization sequence (aa 113-133), one SAND domain (aa 181-280), and two PHD zinc finger domains (aa 299-340 and aa 434-475). Alternate splicing of human AIRE generates isoforms that lack the HSR and SAND domains and/or the second PHD domain. Within aa 476-552, human AIRE shares 65% and 63% aa sequence identity with mouse and rat AIRE, respectively.