Overview

  • Product name
    Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free
    See all ATG9A primary antibodies
  • Description
    Rabbit monoclonal [EPR2450(2)] to ATG9A - BSA and Azide free
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IHC-P, Flow Cyt, IP, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to a region within Human ATG9A.

  • Positive control
    • HepG2, 293T, A375, cell line lysates Mouse brain and rat brain cell lysates Paraffin-embedded human colon tissue
  • General notes

    The formulation and the concentration of this product is compatible for metal-conjugation for mass cytometry (CyTOF®).

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab223528 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 94 kDa.
ICC/IF Use at an assay dependent concentration.

Target

  • Function
    Involved 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 similarities
    Belongs to the ATG9 family.
  • Cellular localization
    Cytoplasmic 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.
  • Information by UniProt
  • Database links
  • Alternative names
    • APG9 autophagy 9-like 1 antibody
    • APG9 like 1 antibody
    • APG9-like 1 antibody
    • APG9L1 antibody
    • ATG9 antibody
    • ATG9 autophagy related 9 homolog A antibody
    • ATG9 autophagy related 9 homolog A (S. cerevisiae) antibody
    • ATG9A antibody
    • ATG9A_HUMAN antibody
    • Autophagy 9-like 1 protein antibody
    • Autophagy related 9A antibody
    • Autophagy related protein 9A antibody
    • Autophagy-related protein 9A antibody
    • mATG9 antibody
    • MGD3208 antibody
    • OTTHUMP00000206046 antibody
    • OTTHUMP00000206048 antibody
    • OTTHUMP00000206049 antibody
    • OTTHUMP00000206062 antibody
    see all

Images

  • Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ATG9A with Purified ab108338 at 1:100 dilution. Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) was used as the secondary antibody at 1:1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

  • This western blot data was generated using the same anti-ATG9A clone (EPR2450(2)] in a different buffer formulation (cat# ab108338).

    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.

  • ab108338 (purified) at 1:20 dilution (2μg) immunoprecipitating ATG9A in HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate.

    Lane 1 (input): HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate 10μg
    Lane 2 (+): ab108338 & HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab108338 in HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution. No band in input lane is due to the boiled lysates
    Blocking and diluting buffer: 5% NFDM/TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

  • Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ATG9A with purified ab108338 at 1:20 dilution (10 µg/ml) (red). Cells were fixed with 100% Methanol and permeabilised with 0.1% Tween-20. A Goat anti rabbit IgG (Alexa Fluor® 488) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human thyroid carcinoma tissue sections labeling ATG9A with Purified ab108338 at 1:50 dilution (4.12 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used at 1:0 dilution. PBS instead of the primary antibody was used as the negative control.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

  • Unpurified 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)

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

  • Unpurified ab108338, at 1/100, staining ATG9A in paraffin-embedded Human colon tissue by Immunohistochemistry.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

  • Unpurified ab108338 at 1/50 dilution, staining ATG9A in HepG2 cells by Immunofluorescence.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).

References

This product has been referenced in:
  • Wu X  et al. Autophagy regulates Notch degradation and modulates stem cell development and neurogenesis. Nat Commun 7:10533 (2016). ICC/IF ; Human . Read more (PubMed: 26837467) »
  • Moreau K  et al. Methods to analyze SNARE-dependent vesicular fusion events that regulate autophagosome biogenesis. Methods 75:19-24 (2015). Read more (PubMed: 25461811) »
See all 7 Publications for this product

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