1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
1/50 - 1/100.
FunctionInvolved in autophagy and cytoplasm to vacuole transport (Cvt) vesicle formation. Plays a key role in the organization of the preautophagosomal structure/phagophore assembly site (PAS), the nucleating site for formation of the sequestering vesicle. Cycles between a juxta-nuclear trans-Golgi network compartment and late endosomes. Nutrient starvation induces accumulation on autophagosomes. Starvation-dependent trafficking requires ULK1, ATG13 and SUPT20H.
Sequence similaritiesBelongs to the ATG9 family.
Cellular localizationCytoplasmic vesicle, autophagosome membrane. Golgi apparatus, trans-Golgi network membrane. Late endosome membrane. Endoplasmic reticulum membrane. Under amino acid starvation or rapamycin treatment, redistributes from a juxtanuclear clustered pool to a dispersed peripheral cytosolic pool. The starvation-induced redistribution depends on ULK1, ATG13, as well as SH3GLB1.
ab108338 staining ATG9A in the human cell line HepG2 (human hepatocellular carcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde and the sample was incubated with the primary antibody at a dilution of 1/40. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)
Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
Predicted band size : 94 kDa
Lane 1: Wild-type HAP1 cell lysate (20 µg) Lane 2: ATG9A knockout HAP1 cell lysate (20 µg) Lane 3: HepG2 cell lysate (20 µg) Lane 4: A375 cell lysate (20 µg) Lanes 1 - 4: Merged signal (red and green). Green - ab108338 observed at 100 and 130 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab108338 was shown to specifically react with ATG9A when ATG9A knockout samples were used. Wild-type and ATG9A knockout samples were subjected to SDS-PAGE. ab108338 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10,000 dilution for 1 h at room temperature before imaging.
Western blot - ATG9A antibody [EPR2450(2)] (ab108338)
All lanes : Anti-ATG9A antibody [EPR2450(2)] (ab108338) at 1/1000 dilution
Lane 1 : HepG2 cell lysate Lane 2 : 293T cell lysate Lane 3 : A375 cell lysate Lane 4 : Mouse brain cell lysate Lane 5 : Rat brain cell lysate
ab108338, at 1/50, staining ATG9A in HepG2 cells by Immunofluorescence.
References for Anti-ATG9A antibody [EPR2450(2)] (ab108338)
This product has been referenced in:
Moreau K et al. Methods to analyze SNARE-dependent vesicular fusion events that regulate autophagosome biogenesis. Methods75:19-24 (2015).
Read more (PubMed: 25461811) »
Zavodszky E et al. Mutation in VPS35 associated with Parkinson's disease impairs WASH complex association and inhibits autophagy. Nat Commun5:3828 (2014).
Read more (PubMed: 24819384) »