Cell Signaling Technology

Product Pathways - Apoptosis

Atg12 Antibody (Human Specific) #2010

Applications Reactivity Sensitivity MW (kDa) Source
W IP IF-IC H Endogenous 16, 55 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Atg12 Antibody (Human Specific) detects endogenous levels of total free and Atg5 bound Atg12 protein.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to residues near the amino terminus of human Atg12. Antibodies are purified by peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Atg12 Antibody (Human Specific).

IF-IC

IF-IC

Confocal immunofluorescent analysis of HCT-116 cells, untreated (left) or chloroquine-treated (right), using Atg12 Antibody (Human Specific) (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

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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