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Cell Biology Proteolysis / Ubiquitin Proteasome / Ubiquitin Ub-like Proteins

Anti-ATG9A antibody [EPR2450(2)] (ab108338)

Price and availability

367 686 ₸

Availability

Order now and get it on Tuesday March 02, 2021

Anti-ATG9A antibody [EPR2450(2)] (ab108338)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)
  • ChIP - Anti-Histone H3 antibody - Nuclear Loading Control and ChIP Grade (ab1791)

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
  • Suitable for: Flow Cyt, WB, IP, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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Overview

  • Product name

    Anti-ATG9A antibody [EPR2450(2)]
    See all ATG9A primary antibodies
  • Description

    Rabbit monoclonal [EPR2450(2)] to ATG9A
  • Host species

    Rabbit
  • Specificity

    The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Mouse
    Rat
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide corresponding to a region within Human ATG9A.

  • Positive control

    • WB: HepG2, 293T, A375, cell line lysates; Mouse brain and rat brain cell lysates IHC-P: Paraffin-embedded human colon tissue; Human thyroid carcinoma tissue. ICC/IF: HepG2 cells.
  • General notes

    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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR2450(2)
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Proteolysis / Ubiquitin
    • Proteasome / Ubiquitin
    • Ub-like Proteins
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Other
    • Cancer
    • Oncoproteins/suppressors
    • Tumor suppressors
    • Other
    • Cardiovascular
    • Heart
    • Autophagy
    • APG gene products
    • Cancer
    • Signal transduction
    • Autophagy
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Autophagy and mitophagy
    • APG gene products
    • Cancer
    • Cell Death
    • Autophagy
    • Signal Transduction
    • Cancer
    • Cell Death
    • Autophagy
    • APG gene products

Images

  • Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Western blot - Anti-ATG9A antibody [EPR2450(2)] (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.

    Unpurified 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.

  • Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] (ab108338) De Pace, R. et al PLoS Genet. 2018 Apr 26;14(4):e1007363. doi: 10.1371/journal.pgen.1007363. eCollection 2018 Apr Reproduced under the Creative Commons license https://creativecommons.org/publicdomain/zero/1.0/

    Accumulation of ATG9A at the TGN of AP-4 μ4 mutant patient fibroblasts.

    Skin fibroblasts were from one control individual and two patients homozygous for mutations in the AP4M1 gene encoding AP-4 μ4.

    Co-immunostaining for endogenous ATG9A (green) and AP-4 ε (red) (B) or GM130 of the fibroblasts.

    ATG9A is detected using ab108338.

     

    (From Figure 4B of De Pace et al)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] (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.

  • Flow Cytometry - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Flow Cytometry - Anti-ATG9A antibody [EPR2450(2)] (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).

  • Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] (ab108338)

    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.

  • Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    All lanes : Anti-ATG9A antibody [EPR2450(2)] (ab108338) at 1/10000 dilution (purified)

    Lane 1 : 293 (Human embryonic kidney epithelial cell) whole cell lysate prepared in non-boiled method
    Lane 2 : 293 (Human embryonic kidney epithelial cell) whole cell lysate prepared in boiled method
    Lane 3 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared in non-boiled method
    Lane 4 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared in boiled method

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 94 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?



    We suggest not to boil the sample after lysis.
    Blocking and diluting buffer: 5% NFDM/TBST

    Exposure time:  
    Left image: 5 seconds
    Right image: 2 seconds

     

  • Immunoprecipitation - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Immunoprecipitation - Anti-ATG9A antibody [EPR2450(2)] (ab108338)

    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.

  • Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Anti-ATG9A antibody [EPR2450(2)] (ab108338) at 1/2000 dilution (purified) + Mouse spinal cord lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 94 kDa



    Blocking and diluting buffer: 5% NFDM/TBST. 

    The lysates are boiled.

  • Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Anti-ATG9A antibody [EPR2450(2)] (ab108338) at 1/2000 dilution (purified) + Rat brain lysates at 15 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 94 kDa



    Blocking and diluting buffer: 5% NFDM/TBST.

    The lysates are boiled.

  • Flow Cytometry - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Flow Cytometry - Anti-ATG9A antibody [EPR2450(2)] (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)

  • Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    All lanes : Anti-ATG9A antibody [EPR2450(2)] (ab108338) at 1/1000 dilution (unpurified)

    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

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 94 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] (ab108338)

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

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

  • Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] (ab108338)

    Unpurified ab108338 at 1/50 dilution, staining ATG9A in HepG2 (Human hepatocellular carcinoma epithelial cell) cells by Immunofluorescence.

  • Anti-ATG9A antibody [EPR2450(2)] (ab108338)
    Anti-ATG9A antibody [EPR2450(2)] (ab108338)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

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