Anti-ATG5 (C-terminal) antibody produced in rabbit

Immunogen

synthetic peptide corresponding to amino acids 262-275 of mouse Atg5, conjugated to KLH via an N-terminal cysteine residue. The corresponding sequence differs by one amino acid in human.

General description

Macroautophagy, usually referred to as autophagy, is a major pathway for bulk degradation of cytoplasmic constituents and organelles. Autophagy is an evolutionary conserved pathway seen in all eukaryotic cells. At least 16 ATG genes, required for autophagosome formation were identified in yeast by genetic screens. For many of these genes, related homologs have been identified in mammals. Atg5, also known as Apg5, is a 32 kDa protein essential for autophagy. Atg5 is covalently modified by Atg12, a ubiquitin-like modifier. This conjugation reaction requires ATP and two enzymes, Atg7 and Atg10, which are E1- and E2-like enzymes, respectively. The Atg12-Atg5 conjugate interacts non-covalently with Atg16. The Atg12-Atg5-Atg16 complex localizes to autophagosome precursors and plays an essential role in autophagosome formation. It has been reported that in addition to the role of Atg5 in the formation of autophagosomes, an Atg5 fragment produced by calpain cleavage has pro-apoptotic properties.

Rabbit polyclonal anti-ATG5 (C-terminal) antibody recognizes mouse and human Atg5-Atg12 complex by immunoblotting, ~56 kDa. Detection of the Atg5-Atg12 complex band by immunoblotting is specifically inhibited with the immunizing peptide.

Antigen Background

ATG5 was found localized to double FYVE domain containing protein 1 (DFCP1)- specific omegasomes. ¹

Physical form

Solution in 0.01 M phos­phate buffered saline, pH 7.4, containing 15 mM sodium azide.

Application

Rabbit polyclonal anti-ATG5 (C-terminal) antibody is used to tag Atg5-Atg12 complex for detection and quantitation by immunocytochemical and immunohistochemical (IHC) techniques. It is used as a probe to determine the presence and roles of Atg5-Atg12 complex in the formation and function of autophagosomes. Rabbit anti-ATG5 was used as primary antibody for immunocytochemistry on HEK293 and Cos7 cells. Cells were fixed in 3.7% formaldehyde in 200 mM Hepes prior to antibody treatment.