		Technical Data
A3935-70	ATG14 (Autophagy-related Protein 14-like Protein, Atg14L, Beclin 1-associated Autophagy-related Key Regulator, BARKOR, KIAA0831, MGC126291, MGC126292)

Description:
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 Atg4 inhibits starvation-induced autophagy (11,12).

Applications:
Suitable for use in Western Blot and Immunoprecipitation. Other applications not tested.

Recommended Dilution:
Western Blot: 1:1000 Incubate membrane with diluted antibody in 5% BSA, 1X TBS, 0.1% Tween-20 at 4°C with gentle shaking, overnight.
Immunoprecipitation: 1:100
Optimal dilutions to be determined by the researcher.

Storage and Stability:
May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Aliquots are stable for at least 12 months. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.

Type	Isotype	Clone	Grade
Pab	IgG		Affinity Purified

Size		Storage	Shipping	Source	Host
100ul		-20°C	Blue Ice	Human	Rabbit

Concentration:

Not determined

Immunogen:

Synthetic peptide corresponding to a region surrounding Val215 of human Atg14.

Purity:

Purified by Protein A affinity chromatography.

Form

Supplied as a liquid in 10mM sodium HEPES, pH 7.5, 150mM sodium chloride, 100ug/ml BSA, 50% glycerol.

Specificity:

endogenous levels of total Atg14. Species Crossreactivity: monkey, mouse, rat.

Intended for research use only. Not for use in human, therapeutic, or diagnostic applications.

(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.