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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

Submit a review Submit a question References (7)
Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
  • Western blot - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
  • Immunoprecipitation - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
  • Flow Cytometry (Intracellular) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
  • Flow Cytometry (Intracellular) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
  • Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
  • Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR2450(2)] to ATG9A - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), IHC-P, IP, WB, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Unconjugated

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: Flow Cyt (Intra), IHC-P, IP, WB, ICC/IFmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

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

  • General notes

    ab223528 is the carrier-free version of ab108338.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    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.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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

    see all

  • 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 - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
    Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

    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).

  • Western blot - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
    Western blot - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

    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.

  • Immunoprecipitation - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
    Immunoprecipitation - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

    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 (Intracellular) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
    Flow Cytometry (Intracellular) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

    Intracellular Flow Cytometry analysis of HepG2 (Human hepatocellular carcinoma epithelial cell) cells labeling ATG9A with purified ab108338 at 1/20 dilution (10µg/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) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

    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).

  • Flow Cytometry (Intracellular) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
    Flow Cytometry (Intracellular) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

    Unpurified ab108338 staining ATG9A in the human cell line HepG2 (human hepatocellular carcinoma) by intracellular 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). 

     

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

    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).

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
    Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

    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).

  • Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)
    Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free (ab223528)

References (7)

Publishing research using ab223528? Please let us know so that we can cite the reference in this datasheet.

ab223528 has been referenced in 7 publications.

  • Wu X  et al. Autophagy regulates Notch degradation and modulates stem cell development and neurogenesis. Nat Commun 7:10533 (2016). ICC/IF ; Human . PubMed: 26837467
  • Moreau K  et al. Methods to analyze SNARE-dependent vesicular fusion events that regulate autophagosome biogenesis. Methods 75:19-24 (2015). PubMed: 25461811
  • Uemura T  et al. A cluster of thin tubular structures mediates transformation of the endoplasmic reticulum to autophagic isolation membrane. Mol Cell Biol 34:1695-706 (2014). PubMed: 24591649
  • Mitzel DN  et al. Age-enhanced endoplasmic reticulum stress contributes to increased Atg9A inhibition of STING-mediated IFN-ß production during Streptococcus pneumoniae infection. J Immunol 192:4273-83 (2014). PubMed: 24670807
  • Zavodszky E  et al. Mutation in VPS35 associated with Parkinson's disease impairs WASH complex association and inhibits autophagy. Nat Commun 5:3828 (2014). ICC/IF ; Human . PubMed: 24819384
  • Moreau K  et al. PICALM modulates autophagy activity and tau accumulation. Nat Commun 5:4998 (2014). ICC/IF ; Human . PubMed: 25241929
  • Puri C  et al. Diverse autophagosome membrane sources coalesce in recycling endosomes. Cell 154:1285-99 (2013). PubMed: 24034251

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