| Product Includes | Product # | Quantity | Mol. Wt | Isotype/Source |
|---|---|---|---|---|
| PI3 Kinase Class III (D9A5) Rabbit mAb | 4263 | 20 µl | 100 kDa | Rabbit |
| PIK3R4 Antibody | 14580 | 20 µl | 153 kDa | Rabbit |
| Beclin-1 (D40C5) Rabbit mAb | 3495 | 20 µl | 60 kDa | Rabbit IgG |
| Atg14 Antibody | 5504 | 20 µl | 65 kDa | Rabbit |
| UVRAG (D2Q1Z) Rabbit mAb | 13115 | 20 µl | 90 kDa | Rabbit IgG |
| Rubicon (D9F7) Rabbit mAb | 8465 | 20 µl | 130 kDa | Rabbit IgG |
| Bif-1 Antibody | 4467 | 20 µl | 42 kDa | Rabbit |
| Atg9A (D4O9D) Rabbit mAb | 13509 | 20 µl | 100-110 kDa | Rabbit IgG |
| Anti-rabbit IgG, HRP-linked Antibody | 7074 | 100 µl | Goat |
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Description
The Autophagy Vesicle Nucleation Antibody Sampler Kit provides an economical means of detecting target proteins involved in autophagosome formation and maturation. The kit contains enough antibody to perform two western blot experiments per primary antibody.
Storage
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 PIK3R4/PI3K class III complex interacts with Beclin-1 to play a role during several stages of autophagy. Autophagosome formation is stimulated when Atg14 complexes with PIK3R4, PI3K class III, and Beclin-1. The UVRAG protein competes with Atg14 for Beclin-1 binding, forming a mutually exclusive complex with PIK3R4, PI3K class III, and Beclin-1 that regulates autophagosome maturation. Autophagosome maturation is impaired in the presence of the Beclin-1-binding protein Rubicon (4,5). Co-expression of PIK3R4 is required for PI3K class III activation and regulation by both Beclin-1/UVRAG and nutrient levels (6). Bif-1 directly binds to UVRAG, forming a complex with Beclin-1, resulting in increased PI3-kinase class III/Vps34 activity required for autophagosome maturation (7). Inhibition of GSK-3β, as seen during nutrient deprivation, results in increased expression of Bif-1, and can contribute to autophagic cell death (8). Atg9A is an integral membrane protein that is required for both the initiation and the expansion of the autophagosome (9,10). Recruitment of Atg9A to the autophagosomal membrane is dynamic and transient as Atg9A also cycles between autophagy-related structures known as omegasomes, the trans-Golgi network (TGN), and endosomes, and at no point becomes a stable component of the autophagosomal membrane (9,11). The precise regulation of Atg9A trafficking is not fully clarified, yet it is suggested to involve p38 mitogen-activated protein kinase (MAPK)-binding protein p38IP and the Beclin-1-binding protein Bif-1 (12,13).
- 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.
- Zhong, Y. et al. (2009) Nat Cell Biol 11, 468-76.
- Sun, Q. et al. (2008) Proc Natl Acad Sci U S A 105, 19211-6.
- Yan, Y. et al. (2009) Biochem J 417, 747-55.
- Takahashi, Y. et al. (2007) Nat Cell Biol 9, 1142-51.
- Yang, J. et al. (2010) J Cell Sci 123, 861-70.
- Young, A.R. et al. (2006) J Cell Sci 119, 3888-900.
- Yamada, T. et al. (2005) J Biol Chem 280, 18283-90.
- Orsi, A. et al. (2012) Mol Biol Cell 23, 1860-73.
- Webber, J.L. and Tooze, S.A. (2010) EMBO J 29, 27-40.
Background References
Trademarks and Patents
Limited Uses
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