Atg16L1 (D6D5) Rabbit mAb

8089:

Immunofluorescence* , Immunoprecipitation , Western Blotting

* Product-specific protocol.

Atg16L1 (D6D5) Rabbit mAb recognizes endogenous levels of total Atg16L1 protein. A background band is detected at 40 kDa in some cell lines.

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Atg16L1 protein.

Western Blotting

Western blot analysis of extracts from various cell lines using Atg16L1 (D6D5) Rabbit mAb.

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with either a myc-tagged human Atg16L1β construct (hAtg16L1β-Myc; +) or a mouse Atg16L1α construct (mAtg16L1α; +), using Atg16L1 (D6D5) Rabbit mAb. The myc-tagged human Atg16L1β construct was kindly provided by Dr. Qing Zhong, University of California Berkeley.

IF-IC

Confocal immunofluorescent analysis of EBSS-starved PANC-1 cells using Atg16L1 (D6D5) Rabbit mAb (green). Blue pseudocolor = DRAQ5^® #4084 (fluorescent DNA dye).

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents. Control of autophagy was largely discovered in yeast and involves proteins encoded by a set of autophagy-related genes (Atg) (1). Formation of autophagic vesicles requires a pair of essential ubiquitin-like conjugation systems, Atg12-Atg5 and Atg8 (LC3)-phosphatidylethanolamine (LC3-PE), which are widely conserved in eukaryotes (2).Mammalian Atg16L1, containing an amino-terminal coiled coil domain and carboxyl-terminal WD-repeats, has multiple isoforms produced by alternative splicing (3,4). Atg16L1 provides a functional link between the two crucial ubiquitin-like conjugation systems of autophagy. Atg16L1 binds Atg5 of the Atg12-Atg5 conjugate forming an 800 kDa multimeric complex (3). The Atg12-Atg-5-Atg16L1 complex localizes to pre-autophagosomal membranes where it determines the site of LC3 lipidation and catalyzes the reaction required for the formation of mature autophagosomes (3,5). Genome-wide association scanning revealed variations in the Atg16L1 gene associated with Crohn's disease (6,7). Mice lacking the coiled coil domain of Atg16L1 have impaired autophagosome formation and elevated inflammatory cytokines, consistent with its role in inflammatory disease pathogenesis (8). Hypomorphic Atg16L1 mice also show defects in autophagy and abnormalities in intestinal Paneth cell function similar to that found in Crohn's disease (9).

1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
2. Ohsumi, Y. (2001) Nat Rev Mol Cell Biol 2, 211-6.
3. Mizushima, N. et al. (2003) J Cell Sci 116, 1679-88.
4. Zheng, H. et al. (2004) DNA Seq 15, 303-5.
5. Fujita, N. et al. (2008) Mol Biol Cell 19, 2092-100.
6. Hampe, J. et al. (2007) Nat Genet 39, 207-11.
7. Rioux, J.D. et al. (2007) Nat Genet 39, 596-604.
8. Saitoh, T. et al. (2008) Nature 456, 264-8.
9. Cadwell, K. et al. (2008) Nature 456, 259-63.

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For Research Use Only. Not For Use In Diagnostic Procedures.