Mouse Monoclonal ATG4A Antibody

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     Western blot analysis of lysate from K562 cell line,  using ATG4A Antibody(Cat.  #P32978). P32978 was diluted at 1:500.  A goat anti-mouse IgG H&L(HRP) at 1:10000 dilution was used as the secondary antibody. Lysate at 20μg.    

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     Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human cervical epithelial adenocarcinoma cell line) cells labeling ATG4A with P32978 at 1/25 dilution, followed by Dylight® 488-conjugated goat anti-mouse IgG   secondary antibody at 1/200 dilution (green). Immunofluorescence image showing cytoplasm staining on HeLa cell line. Cytoplasmic actin is detected with Dylight® 554 Phalloidin  at 1/100 dilution (red).The nuclear counter stain is DAPI (blue).    

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     Overlay histogram showing Hela cells stained with P32978 (green line).  The cells were fixed with 2% paraformaldehyde (10 min) and then permeabilized with 90% methanol for 10 min.  The cells were then icubated in 2% bovine serum albumin to block non-specific protein-protein interactions followed by the antibody (P32978,  1:25 dilution) for 60 min at 37ºC.  The secondary antibody used was Goat-Anti-Mouse IgG, DyLight® 488 Conjugated Highly Cross-Adsorbed(NA168821)) at 1/400 dilution for 40 min at 37ºC.  Isotype control antibody (blue line) was mouse IgG2b (1μg/1x10^6 cells) used under the same conditions.  Acquisition of >10, 000 events was performed.    

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     P32978 staining ATG4A in human brain sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed,  paraffin-embedded sections).  Tissue was fixed with formaldehyde and blocked with 3% BSA for 0. 5 hour at room temperature; antigen retrieval was by heat mediation with a citrate buffer (pH6).  Samples were incubated with primary antibody (1/25) for 1 hours at 37°C.  A undiluted biotinylated goat polyvalent antibody was used as the secondary antibody.    

The ATG4A antibody is a crucial tool for studying autophagy, a cellular degradation process essential for maintaining homeostasis. ATG4A (Autophagy-related protein 4A) is a cysteine protease belonging to the ATG4 family, which plays a pivotal role in autophagosome formation. Specifically, ATG4A processes pro-LC3 (Microtubule-associated protein 1A/1B-light chain 3) by cleaving its C-terminal glycine to generate LC3-I, a cytosolic form that conjugates with phosphatidylethanolamine (PE) to form LC3-II—a key marker of autophagosome membranes. This proteolytic activity is reversible, as ATG4A also delipidates LC3-II to recycle LC3 during autophagy flux. Dysregulation of ATG4A is linked to diseases such as cancer, neurodegenerative disorders, and metabolic conditions, making it a target for therapeutic research.

ATG4A antibodies are widely used in techniques like Western blotting, immunofluorescence, and immunohistochemistry to detect protein expression, localization, and activity in cells or tissues. They help elucidate autophagy mechanisms under stress, nutrient deprivation, or pathological conditions. Commercial ATG4A antibodies are typically raised against specific epitopes (e.g., human ATG4A N-terminal or catalytic domains) and validated for cross-reactivity in model organisms like mice or rats. Researchers rely on these antibodies to investigate ATG4A’s interactions with substrates (e.g., LC3/GABARAP proteins) or regulators (e.g., ATG3. ATG7) and to assess its role in disease progression or treatment responses. Proper validation via knockout controls is critical to ensure specificity in experiments.