Cell Signaling Technology

Product Pathways - Apoptosis

Atg14 Antibody #5504

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R (Mk) Endogenous 65 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Atg14 Antibody detects endogenous levels of total Atg14 protein.

Source / Purification

Polyclonal antibodies were prepared from animals immunized with a synthetic peptide corresponding to a region surrounding Val215 of human Atg14. Antibodies were purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® Atg14 siRNA I #6286 (+) or SignalSilence® Atg14 siRNA II #6287 (+), using Atg14 Antibody #5504 (upper) or β-Tubulin (9F3) Rabbit mAb #2128 (lower). The Atg14 Antibody confirms silencing of Atg14 expression, while the β-Tubulin (9F3) Rabbit mAb is used as a loading control.

Western Blotting

Western Blotting

Western blot analysis of extract from various cell lines using Atg14 Antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a GFP-Atg14 construct (+), using Atg14 Antibody. GFP-Atg14 construct was kindly provided by Dr. Qing Zhong, University of California, Berkeley CA.


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, which are 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 Vps34, including p105/Vps15, Beclin-1, UVRAG, Atg14, and Rubicon (6-12). Atg14L 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, Atg14L localizes to autophagosomes, isolation membranes, and ER and can enhance Vps34 activity. Knockdown of Atg14L inhibits starvation-induced autophagy (11,12).

  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 USA 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.

Application References

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

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