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RabMAb

Anti-ATG9A 抗体 [EPR2450(2)] (ab108338)

製品の概要

  • 製品名
    Anti-ATG9A antibody [EPR2450(2)]
    ATG9A 一次抗体 製品一覧
  • 製品の詳細
    Rabbit monoclonal [EPR2450(2)] to ATG9A
  • アプリケーション
    適用あり: Flow Cyt, WB, IP, IHC-P, ICCmore details
  • 種交差性
    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to a region within Human ATG9A.

  • ポジティブ・コントロール
    • HepG2, 293T, A375, cell line lysates Mouse brain and rat brain cell lysates Paraffin-embedded human colon tissue
  • 特記事項

    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.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab108338 in the following tested applications.

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

アプリケーション Abreviews 特記事項
Flow Cyt Use at an assay dependent concentration.
WB 1/1000 - 1/10000. Predicted molecular weight: 94 kDa.
IP 1/10 - 1/100.
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC 1/50 - 1/100.

ターゲット情報

  • 機能
    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.
  • 配列類似性
    Belongs to the ATG9 family.
  • 細胞内局在
    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
  • 参照データベース
  • 別名
    • 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

画像

  • 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)

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

    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
    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

     



  • 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.

  • 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

    Lysates/proteins at 10 µg per lane.


    Predicted band size : 94 kDa
  • ab108338, at 1/100, staining ATG9A in paraffin-embedded Human colon tissue by Immunohistochemistry.
  • ab108338, at 1/50, staining ATG9A in HepG2 cells by Immunofluorescence.

参考文献

This product has been referenced in:
  • Zhou C  et al. Regulation of mATG9 trafficking by Src- and ULK1-mediated phosphorylation in basal and starvation-induced autophagy. Cell Res 27:184-201 (2017). Read more (PubMed: 27934868) »
  • Nunes J  et al. ATG9A loss confers resistance to trastuzumab via c-Cbl mediated Her2 degradation. Oncotarget 7:27599-612 (2016). Read more (PubMed: 27050377) »

See all 11 Publications for this product

レビューと Q&A

Application
Western blot
Sample
Cow Cell lysate - whole cell (Lung)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (4-12)
Loading amount
30 µg
Specification
Lung
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
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投稿 Apr 05 2017

Application
Western blot
Sample
Mouse Cell lysate - whole cell (neuroblastoma)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (4-12)
Loading amount
30 µg
Specification
neuroblastoma
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
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投稿 Apr 05 2017

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (kidney)
Permeabilization
Yes - 0.2% triton x100
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
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投稿 Mar 24 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
African green monkey Cell (kidney)
Permeabilization
Yes - 0.2% triton x100
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
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投稿 Mar 24 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Brain)
Permeabilization
Yes - 0.3% triton x100
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde
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投稿 Mar 24 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Kidney)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (4-12)
Loading amount
30 µg
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
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投稿 Mar 08 2017

Application
Western blot
Sample
African green monkey Cell lysate - whole cell (Kidney)
Gel Running Conditions
Non-reduced Non-Denaturing (Native) (4-12)
Loading amount
30 µg
Specification
Kidney
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
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投稿 Mar 06 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (breast cancer cell line)
Gel Running Conditions
Reduced Denaturing (gel 8%)
Loading amount
20 µg
Specification
breast cancer cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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投稿 Nov 24 2015

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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