The Autophagy Vesicle Elongation (Atg12 Conjugation) Antibody Sampler Kit provides an economical means of detecting proteins related to autophagy vesicle elongation pathway. The kit contains enough antibody to perform two western blot experiments per primary antibody.
Specificity / Sensitivity
Atg5 (D5F5U) Rabbit mAb recognizes endogenous levels of total Atg5 protein. This antibody is capable of detecting Atg5 conjugated to Atg12. Atg12 (D88H11) Rabbit mAb detects endogenous levels of total free and Atg5 bound Atg12 protein. Atg16L1 (D6D5) Rabbit mAb recognizes endogenous levels of total Atg16L1 protein. A background band is detected at 40 kDa in some cell lines. Atg7 (D12B11) Rabbit mAb recognizes endogenous levels of total Atg7 protein.
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Leu265 of human Atg5 protein, a synthetic peptide corresponding to residues surrounding Ser36 of human Atg12 protein, a synthetic peptide corresponding to residues near the amino terminus of human Atg16L1 protein, and a synthetic peptide corresponding to residues near the amino terminus of human Atg7 protein.
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). Atg16L1 binds Atg5 of the Atg12-Atg5 conjugate forming an 800 kDa multimeric complex (9). The Atg12-Atg-5-Atg16L1 complex localizes to pre-autophagosomal membranes where it determines the site of LC3 lipidation and catalyzes the reaction required for the formation of mature autophagosomes (9,10).