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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.

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).

100 µl
Human, Mouse, Rat

Application Methods: Immunofluorescence (Immunocytochemistry), Western Blotting

Background: GABAA receptor associated protein (GABARAP) is an Atg8 family protein with a key role in autophagy, which was originally discovered as a protein associated with the GABAA receptor regulating receptor trafficking to the plasma membrane (1). Proteins in this family, including microtubule-associated protein light chain 3 (LC3) and GATE-16 (GABARAPL2), become incorporated into the autophagosomal membranes following autophagic stimuli such as starvation (2). Like the other family members, GABARAP is cleaved at its carboxyl terminus, which leads to conjugation by either of the phospholipids phosphatidylethanolamine or phosphatidylserine (3,4). This processing converts GABARAP from a type I to a type II membrane bound form involved in autophagosome biogenesis. Processing of GABARAP involves cleavage by Atg4 family members (5,6) followed by conjugation by the E1 and E2 like enzymes Atg7 and Atg3 (7,8). GABARAPL1/GEC1, a protein that is highly related to GABARAP, was identified as an estrogen inducible gene, and is also associated with autophagosomes (9-11).

The Autophagy Vesicle Elongation (LC3 Conjugation) Antibody Sampler Kit provides an economical means of detecting target proteins related to autophagy vesicle elongation pathway. The kit contains enough antibody to perform two western blots per primary.