重组Anti-ATG7抗体[EPR6251] (ab133528)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6251] to ATG7
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-ATG7抗体[EPR6251]
参阅全部 ATG7 一抗 -
描述
兔单克隆抗体[EPR6251] to ATG7 -
宿主
Rabbit -
经测试应用
适用于: WB, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human ATG7 aa 1-100. The exact sequence is proprietary.
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阳性对照
- 293T, HepG2 and Jurkat whole cell lysate (ab7899), Rat spleen and kidney tissue lysates, mouse spleen and kidney tissue lysates; HT-29 and HeLa cells.
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常规说明
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 ab52472.
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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR6251 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab133528于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB | (9) |
1/10000 - 1/50000. Predicted molecular weight: 77 kDa.
Use 5% non-fat dry milk + TBST for blocking. |
ICC/IF | (1) |
1/100 - 1/500.
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说明 |
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WB
1/10000 - 1/50000. Predicted molecular weight: 77 kDa. Use 5% non-fat dry milk + TBST for blocking. |
ICC/IF
1/100 - 1/500. |
靶标
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功能
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. 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. Plays also 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. -
组织特异性
Widely expressed, especially in kidney, liver, lymph nodes and bone marrow. -
序列相似性
Belongs to the ATG7 family. -
结构域
The C-terminal part of the protein is essential for the dimerization and interaction with ATG3 and ATG12.
The N-terminal FAP motif (residues 15 to 17) is essential for the formation of the ATG89-PE and ATG5-ATG12 conjugates. -
翻译后修饰
Acetylated by EP300. -
细胞定位
Cytoplasm. Preautophagosomal structure. Localizes also to discrete punctae along the ciliary axoneme and to the base of the ciliary axoneme. - Information by UniProt
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数据库链接
- Entrez Gene: 10533 Human
- Entrez Gene: 74244 Mouse
- Entrez Gene: 312647 Rat
- Omim: 608760 Human
- SwissProt: O95352 Human
- SwissProt: Q9D906 Mouse
- SwissProt: Q641Y5 Rat
- Unigene: 38032 Human
see all -
别名
- 1810013K23Rik antibody
- Apg 7 antibody
- APG7 autophagy 7 like antibody
see all
图片
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All lanes : Anti-ATG7 antibody [EPR6251] (ab133528) at 1/10000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : ATG7 knockout HeLa cell lysate
Lane 3 : HepG2 cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 77 kDa
Observed band size: 75 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-ATG7 antibody [EPR6251] staining at 1/10000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (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 ab283307 (knockout 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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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 Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1/200 (2.5 μg/ml). 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.
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All lanes : Anti-ATG7 antibody [EPR6251] (ab133528) at 1/50000 dilution (purified)
Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate
Lane 2 : Mouse spleen lysate
Lane 3 : HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 4 : Mouse kidney lysate
Lane 5 : Rat kidney lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 77 kDa
Observed band size: 77 kDaBlocking and diluting buffer: 5% NFDM/TBST.
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All lanes : Anti-ATG7 antibody [EPR6251] (ab133528) at 1/10000 dilution (purified)
Lane 1 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate
Lane 2 : Rat spleen lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 77 kDa
Observed band size: 77 kDaBlocking and diluting buffer: 5% NFDM/TBST.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: ATG7 knockout HAP1 cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lane 4: HepG2 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab133528 observed at 77 kDa. Red - loading control, ab8245, observed at 37 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 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 (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.
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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. 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.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (69)
ab133528 被引用在 69 文献中.
- Lin YS et al. IGF-1 as a Potential Therapy for Spinocerebellar Ataxia Type 3. Biomedicines 10:N/A (2022). PubMed: 35203722
- Nuth M et al. Discovery of a potent cytotoxic agent that promotes G2/M phase cell cycle arrest and apoptosis in a malignant human pharyngeal squamous carcinoma cell line. Int J Oncol 60:N/A (2022). PubMed: 35211767
- Zhang Z et al. Neuroprotective Effects of a Cholecystokinin Analogue in the 1-Methyl-4-Phenyl-1,2,3,6-Tetrahydropyridine Parkinson's Disease Mouse Model. Front Neurosci 16:814430 (2022). PubMed: 35368248
- Ma B et al. TSPO Ligands Protect against Neuronal Damage Mediated by LPS-Induced BV-2 Microglia Activation. Oxid Med Cell Longev 2022:5896699 (2022). PubMed: 35401924
- Li L et al. Trehalose Protects Keratinocytes against Ultraviolet B Radiation by Activating Autophagy via Regulating TIMP3 and ATG9A. Oxid Med Cell Longev 2022:9366494 (2022). PubMed: 35450405