重组Anti-ATG3抗体[EPR4801] (ab108251)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR4801] to ATG3
- Suitable for: WB, IHC-P, Flow Cyt (Intra), ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-ATG3抗体[EPR4801]
参阅全部 ATG3 一抗 -
描述
兔单克隆抗体[EPR4801] to ATG3 -
宿主
Rabbit -
经测试应用
适用于: WB, IHC-P, Flow Cyt (Intra), ICC/IFmore details
不适用于: IP -
种属反应性
与反应: Human
预测可用于: Mouse, Rat -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HEK293T, K562, HL-60, HeLa or Jurkat whole cell lysate (ab7899). IHC-P: Human muscle tissue. ICC/IF: HeLa cells. Flow Cyt (intra): HeLa cells.
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常规说明
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. Stable for 12 months at -20°C. -
存储溶液
pH: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.1% BSA, 40% Glycerol (glycerin, glycerine), 9.85% Tris glycine, 50% Tissue culture supernatant -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR4801 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab108251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 | (2) |
1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa).
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IHC-P |
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
Perform antigen retrieval before commencing with IHC staining protocol. |
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Flow Cyt (Intra) |
1/2000.
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ICC/IF | (3) |
1/100 - 1/250.
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说明 |
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WB
1/10000 - 1/50000. Detects a band of approximately 40 kDa (predicted molecular weight: 36 kDa). |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Perform antigen retrieval before commencing with IHC staining protocol. |
Flow Cyt (Intra)
1/2000. |
ICC/IF
1/100 - 1/250. |
靶标
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功能
E2-like enzyme involved in autophagy and mitochondrial homeostasis. Catalyzes the conjugation of ATG8-like proteins (GABARAP, GABARAPL1, GABARAPL2 or MAP1LC3A) to phosphatidylethanolamine (PE). PE-conjugation to ATG8-like proteins is essential for autophagy. Preferred substrate is MAP1LC3A. Also acts as an autocatalytic E2-like enzyme, catalyzing the conjugation of ATG12 to itself, ATG12 conjugation to ATG3 playing a role in mitochondrial homeostasis but not in autophagy. ATG7 (E1-like enzyme) facilitates this reaction by forming an E1-E2 complex with ATG3. -
组织特异性
Widely expressed, with a highest expression in heart, skeletal muscle, kidney, liver and placenta. -
序列相似性
Belongs to the ATG3 family. -
翻译后修饰
Conjugated to ATG12 at Lys-243. ATG12-conjugation plays a role in regulation of mitochondrial homeostasis and cell death, while it is not involved in PE-conjugation to ATG8-like proteins and autophagy. -
细胞定位
Cytoplasm. - Information by UniProt
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数据库链接
- Entrez Gene: 64422 Human
- Entrez Gene: 67841 Mouse
- Entrez Gene: 171415 Rat
- Omim: 609606 Human
- SwissProt: Q9NT62 Human
- SwissProt: Q9CPX6 Mouse
- SwissProt: Q6AZ50 Rat
- Unigene: 477126 Human
see all -
别名
- 2610016C12Rik antibody
- Apg 3 antibody
- APG3 antibody
see all
图片
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All lanes : Anti-ATG3 antibody [EPR4801] (ab108251) at 1/1000 dilution
Lane 1 : Wild-type HEK293T cell lysate
Lane 2 : ATG3 knockout HEK293T cell lysate
Lane 3 : K-562 cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution
Predicted band size: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?Lanes 1-4: Merged signal (red and green). Green - ab108251 observed at 40 kDa. Red - loading control ab7291 observed at 50 kDa.
ab108251 Anti-ATG3 antibody [EPR4801] was shown to specifically react with ATG3 in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266707 (knockout cell lysate ab257363) was used. Wild-type and ATG3 knockout samples were subjected to SDS-PAGE. ab108251 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunofluorescent staining of HeLa cells using ab108251 at 1/100.
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All lanes : Anti-ATG3 antibody [EPR4801] (ab108251) at 1/10000 dilution
Lane 1 : K562 cell lysate
Lane 2 : HL-60 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled goat anti-rabbit
Predicted band size: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted? -
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling ATG3 (red) with ab108251 at a 1/2000 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
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ab108251, at a 1/100 dilution, staining Human ATG3 in muscle, using Immunohistochemistry, Formalin/PFA-fixed paraffin-embedded tissue.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (21)
ab108251 被引用在 21 文献中.
- Zhou Y et al. A novel long noncoding RNA SP100-AS1 induces radioresistance of colorectal cancer via sponging miR-622 and stabilizing ATG3. Cell Death Differ 30:111-124 (2023). PubMed: 35978049
- Phadwal K et al. Metformin ameliorates valve interstitial cell calcification by promoting autophagic flux. Sci Rep 13:21435 (2023). PubMed: 38052777
- 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
- Zhu S et al. Proteomic profiling of plasma exosomes from patients with B-cell acute lymphoblastic leukemia. Sci Rep 12:11975 (2022). PubMed: 35831551
- Phadwal K et al. Metformin protects against vascular calcification through the selective degradation of Runx2 by the p62 autophagy receptor. J Cell Physiol 237:4303-4316 (2022). PubMed: 36166694