Scientific BackgroundATG16 L1 plays an essential role in autophagy. It is a homooligomer ant interacts with ATG5. Part of either the minor and major complexes respectively composed of 4 sets of ATG12-ATG5 and ATG16L1 (400 kDa) or 8 sets of ATG12-ATG5 and ATG16L1 (800 kDa). APG16 L1 localises to the cytoplasm, in particularly to preautophagosomal structure (PAS) where it is involved in the membrane targeting of ATG5. There are 5 named isoforms produced by alternative splicing. Genetic variation within ATG16 L1 are associated with susceptibility to inflammatory bowel disease type 1.
Atg16 L1 detected in cytospun HL60 cells by fluorescence confocal microscopy using antibody R-158-100. HL60 cells were fixed with acetone 90 seconds; primary antibody was diluted 1:100 and incubated for 1 hour, 3 x washes and secondary (Goat anti Rb 1:100 Alexa 546 conjugate) for 30 minutes. Cells stained for Atg16 L1 appear in red. The cells were then counter-stained with Hoechst Dye (blue colour).
Western blot (non-reducing) on Jurkat cell lysate using Rabbit antibody to ATG16 L1: whole serum (R-158-100).
Western blot (non-reduced) on Peripheral Blood Mononuclear Cells (PMBC) lysate using Rabbit antibody to ATG16 L1: whole serum (R-158-100) at a dilution of 1: 100 (ECL).
Specific ReferencesCooney R et al (2009). NOD2 stimulation induces autophagy in dendritic cells influencing bacterial handling and antigen presentation. Nat. Med. Dec 6 2009 Scharl M. et al (2011). Protein tyrosine phosphatase nonreceptor type 2 regulates autophagosome formation in human intestinal cells. Inflamm Bowel Dis. 2011 Oct 10.
General ReferencesZheng H, et al. DNA Seq. 15:303-305 (2004). Clark H.F, et al. Genome Res. 13:2265-2270 (2003).