Rabbit Recombinant Monoclonal ATG7 antibody. Suitable for WB, ICC/IF and reacts with Rat, Human, Mouse samples. Cited in 88 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
WB | ICC/IF | |
---|---|---|
Human | Tested | Tested |
Mouse | Tested | Expected |
Rat | Tested | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/10000 - 1/50000 | Notes Use 5% non-fat dry milk + TBST for blocking. |
Species Human | Dilution info 1/10000 - 1/50000 | Notes Use 5% non-fat dry milk + TBST for blocking. |
Species Mouse | Dilution info 1/10000 - 1/50000 | Notes Use 5% non-fat dry milk + TBST for blocking. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/500 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
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E1-like activating enzyme involved in the 2 ubiquitin-like systems required for cytoplasm to vacuole transport (Cvt) and autophagy. Activates ATG12 for its conjugation with ATG5 as well as the ATG8 family proteins for their conjugation with phosphatidylethanolamine. Both systems are needed for the ATG8 association to Cvt vesicles and autophagosomes membranes. Required for autophagic death induced by caspase-8 inhibition. Facilitates LC3-I lipidation with phosphatidylethanolamine to form LC3-II which is found on autophagosomal membranes (PubMed:34161705). Required for mitophagy which contributes to regulate mitochondrial quantity and quality by eliminating the mitochondria to a basal level to fulfill cellular energy requirements and preventing excess ROS production. Modulates p53/TP53 activity to regulate cell cycle and survival during metabolic stress. Also plays a key role in the maintenance of axonal homeostasis, the prevention of axonal degeneration, the maintenance of hematopoietic stem cells, the formation of Paneth cell granules, as well as in adipose differentiation. Plays a role in regulating the liver clock and glucose metabolism by mediating the autophagic degradation of CRY1 (clock repressor) in a time-dependent manner (By similarity).
APG7L, ATG7, Ubiquitin-like modifier-activating enzyme ATG7, ATG12-activating enzyme E1 ATG7, Autophagy-related protein 7, Ubiquitin-activating enzyme E1-like protein, APG7-like, hAGP7
Rabbit Recombinant Monoclonal ATG7 antibody. Suitable for WB, ICC/IF and reacts with Rat, Human, Mouse samples. Cited in 88 publications.
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
We have had 1 attempt at IHC-P with ab133528 in our own lab. We observed both cytoplasmic and nuclear staining on several tissues (including human stomach, kidney and pancreatic cancer), under our experimental conditions. For IHC-P on human tissues, we would recommend using Anti-ATG7 antibody [EP1759Y] ab52472.
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:
For more information, read more on recombinant antibodies.
ATG7 also known as Autophagy Related 7 is an essential protein involved in the autophagy process. It functions as an E1-like enzyme activating and transferring ubiquitin-like proteins such as ATG12 and LC3. The molecular weight of ATG7 is approximately 78 kDa. It is widely expressed in various tissues although expression levels can differ. One can detect ATG7 using immunoassays like ELISA or antibodies specifically targeting ATG7. Understanding its mechanical role is key to studying cellular homeostasis.
ATG7 plays a significant role in autophagy a cellular degradation pathway critical for cell survival under stress. ATG7 contributes to the formation of autophagosomes by facilitating conjugation of ATG8 family proteins including LC3 to phosphatidylethanolamine. It acts within complexes that regulate cellular energy balance and stress responses ensuring cells maintain their function and integrity. Knockdown of ATG7 can impair autophagic flux highlighting its importance in maintaining cellular processes.
ATG7 is a central player in the autophagy pathway influencing cellular metabolism and turnover. It interacts closely with ATG5 and ATG12 in this pathway to form a conjugation system essential for autophagosome elongation. Additionally ATG7 is involved in the mTOR signaling pathway which regulates nutrient sensing and cellular growth. Interaction with proteins like mTOR allows ATG7 to integrate signals from nutrient availability and stress responses finely tuning the autophagy process.
ATG7 dysfunction has connections to cancer and neurodegenerative diseases like Alzheimer's. Abnormal ATG7 activity disrupts autophagic balance possibly leading to the accumulation of damaged proteins and organelles contributing to disease progression. In cancer altered ATG7 expression may influence tumor survival by affecting cellular stress responses. Proteins such as p53 involved in cell cycle regulation often show association with ATG7-related pathways indicating a complex network influencing disease states. Understanding ATG7's role in these conditions can help explore potential therapeutic strategies.
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We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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False colour image of Western blot: Anti-ATG7 antibody [EPR6251] staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab133528 was shown to bind specifically to ATG7. A band was observed at 75 kDa in wild-type HeLa cell lysates with no signal observed at this size in ATG7 knockout cell line Human ATG7 knockout HeLa cell line ab283307 (knockout cell lysate Human ATG7 knockout HeLa cell lysate ab287353). To generate this image, wild-type and ATG7 knockout HeLa cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.
All lanes: Western blot - Anti-ATG7 antibody [EPR6251] (ab133528) at 1/10000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: ATG7 knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human ATG7 knockout HeLa cell line (Human ATG7 knockout HeLa cell line ab283307)
Lane 3: HepG2 cell lysate at 20 µg
Lane 4: Jurkat cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 77 kDa
Observed band size: 75 kDa
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling ATG7 with purified ab133528 at 1/150 dilution (8.5μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/ml). 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.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-ATG7 antibody [EPR6251] (ab133528) at 1/50000 dilution
Lane 1: Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
Lane 2: Mouse spleen lysate at 20 µg
Lane 3: HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate at 20 µg
Lane 4: Mouse kidney lysate at 20 µg
Lane 5: Rat kidney lysate at 20 µg
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: 47 kDa, 77 kDa
Observed band size: 47 kDa, 77 kDa
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Western blot - Anti-ATG7 antibody [EPR6251] (ab133528) at 1/10000 dilution
Lane 1: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Rat spleen lysate at 20 µg
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: 77 kDa
Observed band size: 77 kDa
ab133528 was shown to specifically react with ATG7 when ATG7 knockout samples were used. Wild-type and Apg7 knockout samples were subjected to SDS-PAGE. ab133528 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted 1/10,000 and 1/2000 respectively 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-ATG7 antibody [EPR6251] (ab133528)
Predicted band size: 77 kDa
Immunocytochemistry/Immunofluorescence analysis of HT-29 (Human colorectal adenocarcinoma cell line) labeling ATG7 with purified ab133528 at 1/500 dilution. Cells were fixed with 100% methanol. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077 Goat anti rabbit IgG (Alexa Fluor® 488) at 1/1000 was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
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