Autophagy Atg8 Family Antibody Sampler Kit #64459
Product Information
Kit Usage Information
Protocols
- 3868: Western Blotting, Immunofluorescence*, Flow
- 4599: Western Blotting, Immunohistochemistry (Paraffin)
- 7074: Western Blotting
- 13733: Western Blotting, Immunofluorescence*
- 14256: Western Blotting
- 14736: Western Blotting
- 26632: Western Blotting, Immunofluorescence*, Flow
Product Description
The Autophagy Atg8 Family Antibody Sampler Kit provides an economical means of detecting each of the Atg8 family members. The kit contains enough primary antibody to perform at least two western blot experiments.
Specificity / Sensitivity
Each antibody in the Autophagy Atg8 Family Antibody Sampler Kit detects endogenous levels of its target protein. Cross-reactivity was not observed between family members.
Source / Purification
Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to the amino terminus of human LC3A, LC3B, LC3C, and GABARAPL1, Arg40 of human GABARAP, and the carboxy terminus of GABARAPL2.
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 it has also been associated with a number of physiological processes, including development, differentiation, neurodegenerative diseases, infection, and cancer (3).
Atg8 is a ubiquitin-like protein that is critical for autophagosome formation. Atg8 is synthesized as a precursor protein that is processed by the cysteine protease Atg4, followed by lipidation with phosphatidylethanolamine (PE) in a ubiqutin-like conjugation pathway involving Atg7 and Atg3 (4). This processing of Atg8, which is described as a conversion from type-I to type-II forms, is frequently described as a marker for autophagy. The type-II form of Atg8 is incorporated into maturing autophagosomes and leads to the recruitment of additional autophagy components, including cargo receptors like SQSTM1/p62. While yeast has a single Atg8 gene, many eukaryotes have at least six orthologs, including three microtubule-associated protein 1 light chain 3 (MAP1LC3/LC3) family members (LC3A, LC3B, and LC3C) and three GABAA receptor associated protein (GABARAP) family members (GABARAP, GABARAPL1/GEC1, and GABARAPL2/GATE-16). While highly conserved, these various family members can have important differences in their post-translational processing, expression profile, and protein interactions including distinct cargo receptor. This complexity within the Atg8 family is critical for selective mechanisms of autophagy that have been reported (5, 6).
Atg8 is a ubiquitin-like protein that is critical for autophagosome formation. Atg8 is synthesized as a precursor protein that is processed by the cysteine protease Atg4, followed by lipidation with phosphatidylethanolamine (PE) in a ubiqutin-like conjugation pathway involving Atg7 and Atg3 (4). This processing of Atg8, which is described as a conversion from type-I to type-II forms, is frequently described as a marker for autophagy. The type-II form of Atg8 is incorporated into maturing autophagosomes and leads to the recruitment of additional autophagy components, including cargo receptors like SQSTM1/p62. While yeast has a single Atg8 gene, many eukaryotes have at least six orthologs, including three microtubule-associated protein 1 light chain 3 (MAP1LC3/LC3) family members (LC3A, LC3B, and LC3C) and three GABAA receptor associated protein (GABARAP) family members (GABARAP, GABARAPL1/GEC1, and GABARAPL2/GATE-16). While highly conserved, these various family members can have important differences in their post-translational processing, expression profile, and protein interactions including distinct cargo receptor. This complexity within the Atg8 family is critical for selective mechanisms of autophagy that have been reported (5, 6).
- 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.
- Ichimura, Y. et al. (2000) Nature 408, 488-92.
- Slobodkin, M.R. and Elazar, Z. (2013) Essays Biochem 55, 51-64.
- Schaaf, M.B. et al. (2016) FASEB J 30, 3961-3978.
限制使用
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