Name: Anti-ATG9A antibody [EPR2450(2)]
SKU: ab108338
Rating: 5 (14 reviews)

Anti-ATG9A antibody [EPR2450(2)] ab108338 is a rabbit monoclonal antibody that is used in ATG9A western blotting, IHC, immunofluorescence and flow cytometry. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Antibody clone EPR2450(2) is the most widely used clone for ATG9A on the market
- Specificity confirmed with ATG9A knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation

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Images

Western blot - Anti-ATG9A antibody [EPR2450(2)] (AB108338), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IHC-P	IP	WB	ICC/IF	Flow Cyt (Intra)
Human	Tested	Tested	Tested	Tested	Tested
Mouse	Expected	Expected	Tested	Expected	Expected
Rat	Expected	Expected	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50	Notes The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. For unpurified use 1/100 - 1/250. Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/10 - 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/50 - 1/100	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/20	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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1 product for Alternative Version

1 product for Alternative Product

Target data

See full target information ATG9A

Function

Phospholipid scramblase involved in autophagy by mediating autophagosomal membrane expansion (PubMed:22456507, PubMed:27510922, PubMed:29437695, PubMed:32513819, PubMed:32610138, PubMed:33106659, PubMed:33468622, PubMed:33850023). Cycles between the preautophagosomal structure/phagophore assembly site (PAS) and the cytoplasmic vesicle pool and supplies membrane for the growing autophagosome (PubMed:16940348, PubMed:22456507, PubMed:33106659). Lipid scramblase activity plays a key role in preautophagosomal structure/phagophore assembly by distributing the phospholipids that arrive through ATG2 (ATG2A or ATG2B) from the cytoplasmic to the luminal leaflet of the bilayer, thereby driving autophagosomal membrane expansion (PubMed:33106659). Also required to supply phosphatidylinositol 4-phosphate to the autophagosome initiation site by recruiting the phosphatidylinositol 4-kinase beta (PI4KB) in a process dependent on ARFIP2, but not ARFIP1 (PubMed:30917996). In addition to autophagy, also plays a role in necrotic cell death (By similarity).

Alternative names

APG9L1, ATG9A, Autophagy-related protein 9A, APG9-like 1, mATG9

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR2450(2)
Purification technique: Affinity purification Protein A
Specificity: The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Stable for 12 months at -20°C

Notes

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 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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

ATG9A also known as APG9-like 1 or ATG9 autophagy related 9A is an important component in the autophagy machinery. This protein has a molecular weight of approximately 93 kDa. It is ubiquitously expressed in various tissues with higher expression seen in the heart and skeletal muscle. Mechanically ATG9A is essential for the trafficking of membranes necessary for autophagosome formation. It actively participates in membrane lipids' delivery from donor organelles which are critical for constructing autophagosomes.

Biological function summary

ATG9A facilitates the recycling of cellular materials through a process called autophagy. This protein acts with other autophagy-related proteins to form a complex essential for autophagosome elongation and closure. The ATG9A protein's dynamic movements between the Golgi apparatus and endosomes ensure proper membrane supply for autophagy. It coordinates with the ULK1 complex and various phospholipid-modifying enzymes which are necessary to regulate autophagic flux.

Pathways

ATG9A plays an integral role in the mTOR signaling and MAPK pathways. Both pathways are essential for cellular responses to stress and nutrient availability. In these pathways ATG9A works closely with proteins like beclin 1 and ATG5. Inhibiting mTOR actively induces autophagy where ATG9A contributes to membrane recruitment and elongation of the autophagosomes facilitating the clearance of damaged organelles and proteins.

Associated diseases and disorders

ATG9A shows a significant link to neurodegenerative diseases such as Parkinson's and Alzheimer's disease. The dysfunction of autophagic pathways due to altered ATG9A function leads to the accumulation of damaged proteins and organelles. This event contributes to the pathogenesis of such neurodegenerative diseases. Moreover ATG9A closely associates with proteins like LAMP2 whose mutations similarly lead to defective autophagic processes resulting in the accumulation of unprocessed cellular debris in these disorders.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

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, Anti-GAPDH antibody [6C5] - 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 Anti-GAPDH antibody [6C5] - Loading Control 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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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: Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
Predicted band size: 94 kDa
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/
Immunocytochemistry/ Immunofluorescence - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
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) 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 Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) 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.
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab108338 in HEK-293 (Human embryonic kidney epithelial cell) whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (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.
All lanes: Immunoprecipitation - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
Predicted band size: 94 kDa
Flow Cytometry (Intracellular) - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
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).
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. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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)
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
All lanes: Western blot - 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 at 15 µg
Lane 2: 293 (Human embryonic kidney epithelial cell) whole cell lysate prepared in boiled method at 15 µg
Lane 3: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared in non-boiled method at 15 µg
Lane 4: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate prepared in boiled method at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 94 kDa
Observed band size: 100 kDa
Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
Blocking and diluting buffer: 5% NFDM/TBST.
The lysates are boiled.
All lanes: Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338) at 1/2000 dilution
All lanes: Mouse spinal cord lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 103 kDa, 107 kDa, 110 kDa, 134 kDa, 137 kDa, 14 kDa, 169 kDa, 190 kDa, 20 kDa, 21 kDa, 23 kDa, 24 kDa, 25 kDa, 28 kDa, 30 kDa, 33 kDa, 42 kDa, 43 kDa, 47 kDa, 51 kDa, 53 kDa, 55 kDa, 74 kDa, 83 kDa, 90 kDa, 94 kDa, 96 kDa, 99 kDa
Observed band size: 103 kDa, 120 kDa, 14 kDa, 150 kDa, 169 kDa, 175 kDa, 18 kDa, 193 kDa, 21 kDa, 28 kDa, 32 kDa, 32-42 kDa, 35 kDa, 42 kDa, 44 kDa, 45 kDa, 47 kDa, 51 kDa, 53 kDa, 55 kDa, 90 kDa
Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
Blocking and diluting buffer: 5% NFDM/TBST.
The lysates are boiled.
All lanes: Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338) at 1/2000 dilution
All lanes: Rat brain lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 42 kDa, 47 kDa, 53 kDa, 76 kDa, 82 kDa, 94 kDa
Observed band size: 50-70 kDa, 53 kDa, 72 kDa
Flow Cytometry (Intracellular) - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
Unpurified ab108338 staining ATG9Ain 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)
Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
All lanes: Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338) at 1/1000 dilution
Lane 1: HepG2 cell lysate at 10 µg
Lane 2: 293T cell lysate at 10 µg
Lane 3: A375 cell lysate at 10 µg
Lane 4: Mouse brain cell lysate at 10 µg
Lane 5: Rat brain cell lysate at 10 µg
Predicted band size: 94 kDa
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)
Unpurified ab108338 at 1/50 dilution, staining ATG9A in HepG2 (Human hepatocellular carcinoma epithelial cell) cells by Immunofluorescence.