Cell Signaling Technology

Product Pathways - Apoptosis

Atg5 (D1G9) Rabbit mAb #8540

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M R Mk (B) (Dg) (Pg) Endogenous 55 Rabbit IgG

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

Protocols

Specificity / Sensitivity

Atg5 (D1G9) Rabbit mAb recognizes endogenous levels of total Atg5 protein. This antibody is capable of detecting Atg5 when conjugated to Atg12.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro187 of human Atg5 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Atg5 (D1G9) Rabbit mAb.

IP

IP

Immunoprecipitation of Atg5 from HT-1080 cells using Atg5 (D1G9) Rabbit mAb. Western blot detection was performed using the same antibody (upper), or with Atg12 (D88H11) Rabbit mAb #4180 (lower). Lane 1 is 10% input, lane 2 is precipitated with non-specific rabbit IgG, lane 3 is precipitated with Atg5 (D1G9) Rabbit mAb.

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. Formation of the autophagosome involves a ubiquitin-like conjugation system in which Atg12 is covalently bound to Atg5 and targeted to autophagosome vesicles (4-6). This conjugation reaction is mediated by the ubiquitin E1-like enzyme Atg7 and the E2-like enzyme Atg10 (7,8).

  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. Mizushima, N. et al. (1998) J Biol Chem 273, 33889-92.
  5. Mizushima, N. et al. (1998) Nature 395, 395-8.
  6. Suzuki, K. et al. (2001) EMBO J 20, 5971-81.
  7. Tanida, I. et al. (1999) Mol Biol Cell 10, 1367-79.
  8. Shintani, T. et al. (1999) EMBO J 18, 5234-41.

Application References

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Companion Products

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

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