Autophagy Induction (ULK1 Complex) Antibody Sampler Kit #46486
Product Information
Kit Usage Information
Protocols
- 5869: Western Blotting, Immunoprecipitation (Agarose)
- 7074: Western Blotting
- 8054: Western Blotting, Immunoprecipitation (Magnetic)
- 12436: Western Blotting, Immunoprecipitation (Magnetic)
- 13273: Western Blotting, Immunoprecipitation (Agarose)
- 13492: Western Blotting
- 14202: Western Blotting, Immunoprecipitation (Agarose), Immunofluorescence, Flow
Product Description
Specificity / Sensitivity
ULK1 (D8H5) Rabbit mAb, Atg13 (D4P1K) Rabbit mAb, FIP200 (D10D11) Rabbit mAb, and Atg101 (E1Z4W) Rabbit mAb recognize total endogenous levels of the corresponding target proteins irrespective of phosphorylation state. Phospho-ULK1 (Ser555) (D1H4) Rabbit mAb detects endogenous levels of ULK1 only when phosphorylated at Ser555 of mouse ULK1 (equivalent to Ser556 of human ULK1). Bands of unknown origin are detected between 90 and 100 kDa. Phospho-ULK1 (Ser757) (D7O6U) Rabbit mAb recognizes endogenous levels of ULK1 protein only when phosphorylated at Ser757 of mouse ULK1 (equivalent to Ser758 of human ULK1).
Source / Purification
Monoclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Arg600 of human ULK1 protein, residues surrounding Asp462 of human Atg13 protein, residues surrounding Val177 of human Atg101 protein, or residues near the carboxy terminus of human FIP200 protein. Phospho-specific monoclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser555 of mouse ULK1 protein (equivalent to Ser556 of human ULK1) or residues surrounding Ser757 of mouse ULK1 protein (equivalent to Ser758 of human ULK1).
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. ULK1, Atg13, and FIP200 form a complex that localizes to autophagic isolation membranes and regulates autophagosome biogenesis (4-6). mTOR phosphorylates both Atg13 and ULK1, suppressing ULK1 kinase activity and autophagy (5-7). Interaction between Atg101 and Atg13 can be important for the stability and basal phosphorylation of Atg13 and ULK1 (8,9). AMPK, activated during low nutrient conditions, directly phosphorylates ULK1 at multiple sites including Ser317, Ser555, and Ser777 (7,10). Conversely, mTOR, which is a regulator of cell growth and is an inhibitor of autophagy, phosphorylates ULK1 at Ser757 and disrupts the interaction between ULK1 and AMPK (7).
- 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.
- 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.
- Mercer, C.A. et al. (2009) Autophagy 5, 649-62.
- Hosokawa, N. et al. (2009) Autophagy 5, 973-9.
- Egan, D.F. et al. (2011) Science 331, 456-61.
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