Cell Signaling Technology

Product Pathways - Autophagy Signaling

Phospho-Atg14 (Ser29) Antibody #13155

No. Size Price
13155S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
13155 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse, Endogenous 65 Rabbit

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting,

Specificity / Sensitivity

Phospho-Atg14 (Ser29) Antibody recognizes endogenous levels of Atg14 protein only when phosphorylated at Ser29. This antibody also cross-reacts with unidentified proteins of 32 kDa, 80 kDa and 100 kDa.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser29 of human Atg14 protein. Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing GFP-tagged human Atg14 protein (hAtg14-GFP; +) or mouse ULK1 protein (mULK1; +), using Phospho-Atg14 (Ser29) Antibody (upper) and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from A172 cells, untreated (-) or starved for 2 hours with Earle's Basic Salt Solution (EBSS; +), using Phospho-Atg14 (Ser29) Antibody (upper), Atg14 Antibody #5504 (middle), and GAPDH (D16H11) XP® Rabbit mAb #5174 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from C2C12 cells, untreated or starved with Earle's Basic Salt Solution (EBSS) for the indicated times using Phospho-Atg14 (Ser29) Antibody (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

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 is also 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 is directed by a number of autophagy-related (Atg) genes. These proteins are involved in the formation of autophagosomes, cytoplasmic vacuoles that are delivered to lysosomes for degradation. The class III type phosphoinositide 3-kinase (PI3K) Vps34 regulates vacuolar trafficking and autophagy (4,5). Multiple proteins associate with Vsp34, including p105/Vsp15, Beclin-1, UVRAG, Atg14, and Rubicon, to determine Vsp34 function (6-12). Atg14 and Rubicon were identified based on their ability to bind to Beclin-1 and participate in unique complexes with opposing functions (9-12). Rubicon, which localizes to the endosome and lysosome, inhibits Vps34 lipid kinase activity; knockdown of Rubicon enhances autophagy and endocytic trafficking (11,12). In contrast, Atg14 localizes to autophagosomes, isolation membranes and ER, and can enhance Vps34 activity. Knockdown of Atg14 inhibits starvation-induced autophagy (11,12).

The serine/threonine kinase ULK1 phosphorylates Atg14 at Ser29 to promote autophagsome formation (13).

  1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot Cell 1, 11-21.
  2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ 12 Suppl 2, 1509-18.
  3. Levine, B. and Yuan, J. (2005) J Clin Invest 115, 2679-88.
  4. Corvera, S. (2001) Traffic 2, 859-66.
  5. Yan, Y. and Backer, J.M. (2007) Biochem Soc Trans 35, 239-41.
  6. Stack, J.H. et al. (1995) J Cell Biol 129, 321-34.
  7. Zeng, X. et al. (2006) J Cell Sci 119, 259-70.
  8. Liang, C. et al. (2006) Nat Cell Biol 8, 688-99.
  9. Itakura, E. et al. (2008) Mol Biol Cell 19, 5360-72.
  10. Sun, Q. et al. (2008) Proc Natl Acad Sci U S A 105, 19211-6.
  11. Zhong, Y. et al. (2009) Nat Cell Biol 11, 468-76.
  12. Matsunaga, K. et al. (2009) Nat Cell Biol 11, 385-96.
  13. Park, J.M. et al. (2016) Autophagy 12, 547-564.

Application References

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