Anti-ATG9A 抗体 [EPR2450(2)]
Anti-ATG9A antibody [EPR2450(2)]
- RabMAb
- Recombinant
- KO Validated
- 了解详情
5
(14 Reviews)
|
(83 Publications)
Anti-ATG9A antibody [EPR2450(2)] (ab108338) is a rabbit monoclonal antibody detecting ATG9A in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, IHC-P, ICC/IF. Suitable for Human, Mouse, Rat.
- KO validated for confirmed specificity
- Biophysical QC for unrivalled batch-batch consistency
- Over 80 publications
查看别名
APG9L1, ATG9A, Autophagy-related protein 9A, APG9-like 1, mATG9
- IHC-P
Lab
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ATG9A antibody [EPR2450(2)] (AB108338)
Immunohistochemical analysis of formalin fixed paraffin embedded human colon labelling ATG9A with ab108338 at a concentration of 0.5µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins. ab108338 anti-ATG9A antibody [EPR2450(2)] was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual).
- WB
Unknown
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
false
- ICC/IF
PubMed
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)
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/
- Flow Cyt (Intra)
Lab
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)
- IHC-P
Unknown
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.
- Flow Cyt (Intra)
Unknown
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).
- IHC-P
Unknown
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.
- ICC/IF
Unknown
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.
- ICC/IF
Unknown
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.
- IP
Lab
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.
All lanes:
Immunoprecipitation - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
Predicted band size: 94 kDa
false
- WB
Lab
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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution
Predicted band size: 94 kDa
Observed band size: 100 kDa
false
- WB
Lab
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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) 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
false
- WB
Lab
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.
All lanes:
Western blot - Anti-ATG9A antibody [EPR2450(2)] (ab108338)
Predicted band size: 94 kDa
false
- WB
Lab
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) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) 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
false
不同偶联物与剂型 (8)
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665 Alexa Fluor® 647
Alexa Fluor® 647 Anti-ATG9A antibody [EPR2450(2)]
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519 Alexa Fluor® 488
Alexa Fluor® 488 Anti-ATG9A antibody [EPR2450(2)]
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Anti-ATG9A antibody [EPR2450(2)] - BSA and Azide free
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660 APC
APC Anti-ATG9A antibody [EPR2450(2)]
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578 PE
PE Anti-ATG9A antibody [EPR2450(2)]
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603 Alexa Fluor® 568
Alexa Fluor® 568 Anti-ATG9A antibody [EPR2450(2)]
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617 Alexa Fluor® 594
Alexa Fluor® 594 Anti-ATG9A antibody [EPR2450(2)]
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775 Alexa Fluor® 750
Alexa Fluor® 750 Anti-ATG9A antibody [EPR2450(2)]
反应性数据
产品详情
What is this antibody validated in?
Anti-ATG9A antibody [EPR2450(2)] (ab108338) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), Immunocytochemistry/immunofluorescence (ICC/IF) in Human, Mouse, Rat samples.
What is the molecular weight of ATG9A?
Anti-ATG9A [EPR2450(2)] (ab108338) specifically detects a band for ATG9A (UniProt: Q7Z3C6) at a molecular weight of 94kDa.
Trusted by the scientific community
Anti-ATG9A [EPR2450(2)] (ab108338) was first used in a scientific publication in 2011 and has been cited over 80 times in peer-reviewed journals.
Reviewed by scientists
Anti-ATG9A [EPR2450(2)] (ab108338) has over 10 independent reviews from customers.
Trial sizes available!
Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies.
Specificity confirmed
The specificity of Anti-ATG9A antibody [EPR2450(2)] (ab108338) has been confirmed by Western blot testing in ATG9A Knockout HAP1 cells.
Other related products
We have a range of other formats of antibody clone [EPR2450(2)] also available for your convenience: ab108338, Alexa Fluor® 488 - ab206252, Alexa Fluor® 647 - ab206253, Carrier free - ab223528, ab309518, Carrier free - ab309519, APC - ab310808, PE - ab310890, Alexa Fluor® 594 - ab311672, Alexa Fluor® 568 - ab312947, Alexa Fluor® 750 - ab320894
Patented technology
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
What are the advantages of a recombinant monoclonal antibody?
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.
性能和储存信息
形式
纯化工艺
存储溶液
运输条件
推荐的短期储存时间
推荐的短期储存条件
推荐的长期储存条件
分装信息
储存信息
补充信息
This supplementary information is collated from multiple sources and compiled automatically.
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.
产品实验方案
- Visit the General protocols
- Visit the Troubleshooting
靶点信息
文献 (83)
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