Rabbit Polyclonal ATG12(N-term) Antibody

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     Anti-APG12L Antibody (E3) at 1:500 dilution + HCT116 whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG,  (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 15 kDa Blocking/Dilution buffer: 5% NFDM/TBST.    

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     All lanes : Anti-APG12L Antibody (E3) at 1:2000 dilution Lane 1: A549 whole cell lysate Lane 2: Hela whole cell lysate Lysates/proteins at 20 µg per lane. Secondary Goat Anti-Rabbit IgG,  (H+L), Peroxidase conjugated at 1/10000 dilution. Predicted band size : 15 kDa Blocking/Dilution buffer: 5% NFDM/TBST.    

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     Fluorescent image of U251 cells stained with ATG12 (N-term) antibody. U251 cells were treated with Chloroquine (50 μM,16h), then fixed with 4% PFA (20 min), permeabilized with Triton X-100 (0.2%, 30 min). Cells were then incubated with P33999 ATG12 (N-term) primary antibody (1:200, 2 h at room temperature). For secondary antibody, Alexa Fluor® 488 conjugated donkey anti-rabbit antibody (green) was used (1:1000, 1h). Nuclei were counterstained with Hoechst 33342 (blue) (10 μg/ml, 5 min). ATG12 immunoreactivity is localized to autophagic vacuoles in the cytoplasm of U251 cells.    

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     Formalin-fixed and paraffin-embedded human cancer tissue reacted with the primary antibody, which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated. BC = breast carcinoma; HC = hepatocarcinoma.    

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     Formalin-fixed and paraffin-embedded human lung carcinoma tissue reacted with Autophagy APG12L antibody (N-term) (Cat.#P33999), which was peroxidase-conjugated to the secondary antibody, followed by DAB staining. This data demonstrates the use of this antibody for immunohistochemistry; clinical relevance has not been evaluated.    

The ATG12 (N-term) antibody is a widely used tool in autophagy research, targeting the N-terminal region of the autophagy-related 12 (ATG12) protein. ATG12 is a critical component of the ubiquitin-like conjugation system essential for autophagosome formation, a key step in macroautophagy. During this process, ATG12 covalently binds to ATG5 via a ubiquitin-like conjugation mechanism, forming the ATG12-ATG5 complex. This complex further interacts with ATG16L1 to create a multimeric complex that promotes LC3 lipidation and autophagosome membrane expansion.

The ATG12 (N-term) antibody specifically detects endogenous full-length ATG12 (~18 kDa) and helps distinguish unprocessed ATG12 from its conjugated form (ATG12-ATG5. ~55 kDa) in techniques like Western blotting, immunofluorescence, or immunohistochemistry. It is particularly valuable for monitoring autophagy flux, as ATG12 expression levels or conjugation status often correlate with autophagic activity. Researchers frequently employ this antibody in studies investigating autophagy's role in cancer, neurodegenerative diseases, infections, and metabolic disorders. Its specificity is validated through knockout controls or siRNA-mediated ATG12 depletion. By enabling precise detection of ATG12 dynamics, this antibody supports mechanistic insights into autophagy regulation and its implications in health and disease.