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Anti-ATG4A Antibody

Rabbit polyclonal antibody to ATG4A

Western blot analysis of ATG4A expression in HCT116 (A), HepG2 (B), CT26 (C), PC12 (D) whole cell lysates.
Immunohistochemical analysis of ATG4A staining in human breast cancer formalin fixed paraffin embedded tissue section. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0). The section was then incubated with the antibody at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
Immunofluorescent analysis of ATG4A staining in A549 cells. Formalin-fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 5-10 minutes and blocked with 3% BSA-PBS for 30 minutes at room temperature. Cells were probed with the primary antibody in 3% BSA-PBS and incubated overnight at 4 °C in a hidified chamber. Cells were washed with PBST and incubated with Alexa Fluor 647-conjugated secondary antibody (red) in PBS at room temperature in the dark.

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Product Information
Application Applications Legend: E=ELISA WB=Western Blotting IHC=Immunohistochemistry IHC-P=Immunohistochemistry (Paraffin) IP=Immunoprecipitation IF=Immunofluorescence IC=Immunochemistry ICC=Immunocytochemistry FC=Flow Cytometry DB=Dot Blot	WB, IF/IC, IHC
Primary Accession	Q8WYN0
Other Accession	Q8C9S8
Reactivity	Human, Mouse, Rat
Host	Rabbit
Clonality	Polyclonal
Calculated MW	45378 Da

Additional Information
Gene ID	115201
Other Names	APG4A; AUTL2; Cysteine protease ATG4A; AUT-like 2 cysteine endopeptidase; Autophagin-2; Autophagy-related cysteine endopeptidase 2; Autophagy-related protein 4 homolog A; hAPG4A
Target/Specificity	Recognizes endogenous levels of ATG4A protein.
Dilution	WB~~WB (1/500 - 1/1000), IHC (1/50 - 1/100), IF/IC (1/100 - 1/500) IF/IC~~N/A IHC~~WB (1/500 - 1/1000), IHC (1/50 - 1/100), IF/IC (1/100 - 1/500)
Format	Liquid in 0.42% Potassium phosphate, 0.87% Sodium chloride, pH 7.3, 30% glycerol, and 0.09% (W/V) sodium azide.
Storage	Store at -20 °C.Stable for 12 months from date of receipt

Protein Information
Name	ATG4A {ECO:0000303\|Ref.20, ECO:0000312\|HGNC:HGNC:16489}
Function	Cysteine protease that plays a key role in autophagy by mediating both proteolytic activation and delipidation of ATG8 family proteins (PubMed:12473658, PubMed:15169837, PubMed:17347651, PubMed:21177865, PubMed:21245471, PubMed:22302004, PubMed:32732290). The protease activity is required for proteolytic activation of ATG8 family proteins: cleaves the C-terminal amino acid of ATG8 proteins to reveal a C-terminal glycine (PubMed:12473658, PubMed:15169837, PubMed:17347651, PubMed:21177865, PubMed:21245471, PubMed:22302004). Exposure of the glycine at the C-terminus is essential for ATG8 proteins conjugation to phosphatidylethanolamine (PE) and insertion to membranes, which is necessary for autophagy (PubMed:12473658, PubMed:15169837, PubMed:17347651, PubMed:21177865, PubMed:21245471, PubMed:22302004). Preferred substrate is GABARAPL2 followed by MAP1LC3A and GABARAP (PubMed:12473658, PubMed:15169837, PubMed:17347651, PubMed:21177865, PubMed:21245471, PubMed:22302004). Protease activity is also required to counteract formation of high-molecular weight conjugates of ATG8 proteins (ATG8ylation): acts as a deubiquitinating- like enzyme that removes ATG8 conjugated to other proteins, such as ATG3 (PubMed:31315929, PubMed:33773106). In addition to the protease activity, also mediates delipidation of ATG8 family proteins (PubMed:29458288, PubMed:33909989). Catalyzes delipidation of PE- conjugated forms of ATG8 proteins during macroautophagy (PubMed:29458288, PubMed:33909989). Compared to ATG4B, the major protein for proteolytic activation of ATG8 proteins, shows weaker ability to cleave the C-terminal amino acid of ATG8 proteins, while it displays stronger delipidation activity (PubMed:29458288). Involved in phagophore growth during mitophagy independently of its protease activity and of ATG8 proteins: acts by regulating ATG9A trafficking to mitochondria and promoting phagophore-endoplasmic reticulum contacts during the lipid transfer phase of mitophagy (PubMed:33773106).
Cellular Location	Cytoplasm {ECO:0000250\|UniProtKB:Q8BGE6}.