Revision 3
#26839
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For Research Use Only. Not for Use in Diagnostic Procedures.
Applications:
W, IP
Reactivity:
H
Sensitivity:
Transfected Only
MW (kDa):
72
Source/Isotype:
Rabbit IgG
UniProt ID:
#O75143
Entrez-Gene Id:
9776
Product Usage Information
Application | Dilution |
---|---|
Western Blotting | 1:1000 |
Immunoprecipitation | 1:50 |
Storage
Specificity/Sensitivity
Phospho-Atg13 (Ser355) (D6J1W) Rabbit mAb is recommended for transfected levels of human Atg13 protein only when phosphorylated at Ser355 (or Ser354 for mouse Atg13). This antibody can weakly detect endogenous levels of phosphorylated Atg13. A band of unknown origin is detected at 25 kDa in some cell lines.
Species predicted to react based on 100% sequence homology
Mouse, Rat
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic phospho-peptide corresponding to residues surrounding Ser355 of human Atg13 protein (Ser318 of isoform 2 of Atg13).
Background
Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation but has also been associated with a number of physiological processes including development, differentiation, neurodegeneration, infection, and cancer (3). The molecular machinery of autophagy was largely discovered in yeast and referred to as autophagy-related (Atg) genes.
Atg13/Apg13 was originally identified in yeast as a constitutively expressed protein that was genetically linked to Atg1/Apg1, a protein kinase required for autophagy (4). Overexpression of Atg1 suppresses the defects in autophagy observed in Atg13 mutants (4). Autophagy requires a direct association between Atg1 and Atg13, and is inhibited by TOR-dependent phosphorylation of Atg13 under high-nutrient conditions (5). Similarly, mammalian Atg13 forms a complex with the Atg1 homologues ULK1/2, along with FIP200, which localizes to autophagic isolation membranes and regulates autophagosome biogenesis (6-8). mTOR phosphorylates both Atg13 and ULK1, suppressing ULK1 kinase activity and autophagy (7-9). ULK1 can directly phosphorylate Atg13 at a yet unidentified site, presumably to promote autophagy (7,8). Additional studies suggest that Atg13 and FIP200 can function independently of ULK1 and ULK2 to induce autophagy through an unknown mechanism (10).
ULK1-dependent phosphorylation of Atg13 at Ser355, which corresponds to Ser318 of isoform 2 of Atg13, leads to the recruitment of Atg13 to damaged mitochondria, enabling efficient mitophagy (11).
Background References
- Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
- Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
- Levine, B. and Yuan, J. (2005) J Clin Invest 115, 2679-88.
- Funakoshi, T. et al. (1997) Gene 192, 207-13.
- Kamada, Y. et al. (2000) J Cell Biol 150, 1507-13.
- Ganley, I.G. et al. (2009) J Biol Chem 284, 12297-305.
- Hosokawa, N. et al. (2009) Mol Biol Cell 20, 1981-91.
- Jung, C.H. et al. (2009) Mol Biol Cell 20, 1992-2003.
- Kim, J. et al. (2011) Nat Cell Biol 13, 132-41.
- Alers, S. et al. (2011) Autophagy 7, 1423-33.
- Joo, J.H. et al. (2011) Mol Cell 43, 572-85.
Species Reactivity
Species reactivity is determined by testing in at least one approved application (e.g., western blot).
Western Blot Buffer
IMPORTANT: For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% Tween® 20 at 4°C with gentle shaking, overnight.
Applications Key
W: Western Blotting IP: Immunoprecipitation
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
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XP is a registered trademark of Cell Signaling Technology, Inc.
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Limited Uses
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Revision 3
Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human Atg13 (hAtg13-Myc/DDK; +) or mouse ULK1 (mULK1; +), using Phospho-Atg13 (Ser355) (D6J1W) Rabbit mAb.
Immunoprecipitation of Phospho-Atg13 (Ser355) from extracts of PANC-1 cells starved with Hank's Balanced Salt Solution (HBSS) (2 hr). Lane 1 is 10% input, lane 2 is Rabbit (DA1E) mAb IgG XP® Isotype Control #3900, and lane 3 is Phospho-Atg13 (Ser355) (D6J1W) Rabbit mAb. Western blot was performed using Phospho-Atg13 (Ser355) (D6J1W) Rabbit mAb.