重组Anti-APG5L/ATG5抗体[EPR1755(2)] (ab108327)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR1755(2)] to APG5L/ATG5
- Suitable for: ICC/IF, WB, IP, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-APG5L/ATG5抗体[EPR1755(2)]
参阅全部 APG5L/ATG5 一抗 -
描述
兔单克隆抗体[EPR1755(2)] to APG5L/ATG5 -
宿主
Rabbit -
经测试应用
适用于: ICC/IF, WB, IP, IHC-Pmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- Raji, HeLa, HT-1080, Human fetal kidney, C6, Raw264.7, PC-12 and NIH3T3 cell lysates; Human hepatocellular carcinoma and Human ovarian adenocarcinoma tissue
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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: 7.2
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
EPR1755(2) -
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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KO cell lines
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Recombinant Protein
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Related Products
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab108327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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ICC/IF | (1) |
1/100 - 1/250.
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WB | (4) |
1/1000 - 1/10000. Predicted molecular weight: 32 kDa.
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IP |
1/10 - 1/100.
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IHC-P | (1) |
1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Antigen retrieval is recommended. |
说明 |
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ICC/IF
1/100 - 1/250. |
WB
1/1000 - 1/10000. Predicted molecular weight: 32 kDa. |
IP
1/10 - 1/100. |
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. Antigen retrieval is recommended. |
靶标
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功能
Involved in autophagic vesicle formation. Conjugation with ATG12, through a ubiquitin-like conjugating system involving ATG7 as an E1-like activating enzyme and ATG10 as an E2-like conjugating enzyme, is essential for its function. The ATG12-ATG5 conjugate acts as an E3-like enzyme which is required for lipidation of ATG8 family proteins and their association to the vesicle membranes. Involved in mitochondrial quality control after oxidative damage, and in subsequent cellular longevity. The ATG12-ATG5 conjugate also negatively regulates the innate antiviral immune response by blocking the type I IFN production pathway through direct association with RARRES3 and MAVS. Also plays a role in translation or delivery of incoming viral RNA to the translation apparatus. Plays a critical role in multiple aspects of lymphocyte development and is essential for both B and T lymphocyte survival and proliferation. Required for optimal processing and presentation of antigens for MHC II. Involved in the maintenance of axon morphology and membrane structures, as well as in normal adipocyte differentiation. Promotes primary ciliogenesis through removal of OFD1 from centriolar satellites and degradation of IFT20 via the autophagic pathway.
May play an important role in the apoptotic process, possibly within the modified cytoskeleton. Its expression is a relatively late event in the apoptotic process, occurring downstream of caspase activity. Plays a crucial role in IFN-gamma-induced autophagic cell death by interacting with FADD. -
组织特异性
Ubiquitous. The mRNA is present at similar levels in viable and apoptotic cells, whereas the protein is dramatically highly expressed in apoptotic cells. -
序列相似性
Belongs to the ATG5 family. -
翻译后修饰
Conjugated to ATG12; which is essential for autophagy, but is not required for association with isolation membrane.
Acetylated by EP300. -
细胞定位
Cytoplasm. Preautophagosomal structure membrane. Colocalizes with nonmuscle actin. The conjugate detaches from the membrane immediately before or after autophagosome formation is completed (By similarity). 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: 9474 Human
- Entrez Gene: 11793 Mouse
- Entrez Gene: 365601 Rat
- Omim: 604261 Human
- SwissProt: Q9H1Y0 Human
- SwissProt: Q99J83 Mouse
- SwissProt: Q3MQ06 Rat
- Unigene: 486063 Human
see all -
别名
- APG 5 antibody
- APG 5L antibody
- APG5 antibody
see all
图片
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All lanes : Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/1000 dilution
Lane 1 : Wild-type THP-1 cell lysate
Lane 2 : ATG5 knockout THP-1 cell lysate
Lane 3 : U-87 MG cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 32 kDa
Observed band size: 50 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-APG5L/ATG5 antibody [EPR1755(2)] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab108327 was shown to bind specifically to APG5L/ATG5. A band was observed at 50 kDa in wild-type THP-1 cell lysates with no signal observed at this size in ATG5 knockout cell line ab277835 (knockout cell lysate ab290722). To generate this image, wild-type and ATG5 knockout THP-1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % 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 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: APG5L/ATG5 knockout HAP1 cell lysate (20 µg)
Lane 3: Raji cell lysate (20 µg)
Lane 4: Jeg-3 cell lysate (20 µg)Lanes 1 - 4: Merged signal (red and green). Green - ab108327 observed at 52 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab108327 was shown to specifically react with APG5L/ATG5 when APG5L/ATG5 knockout samples were used. Wild-type and APG5L/ATG5 knockout samples were subjected to SDS-PAGE. ab108327 and ab8245 (loading control to GAPDH) were both diluted 1/1000 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 1/10000 dilution for 1 h at room temperature before imaging. -
Unpurified ab108327, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human ovarian adenocarcinoma tissue by Immunohistochemistry.
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Immunofluorescence staining of MCF7 cells with purified ab108327 at a working dilution of 1/150, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 100% methanol and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab108327 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.
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All lanes : Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/10000 dilution (purified)
Lane 1 : C6 cell lysate
Lane 2 : PC-12 cell lysate
Lane 3 : NIH/3T3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/10000 dilution (purified) + Raji cell lysate at 20 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/10000 dilution
Predicted band size: 32 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/2000 dilution (purified) + HT-1080 cell lysate at 20 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 55 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/2000 dilution (purified) + human fetal kidney at 10 µg
Secondary
HRP goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 32 kDa
Observed band size: 32,55 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST -
Immunohistochemical staining of paraffin embedded human cervical carcinoma with purified ab180327 at a working dilution of 1/150. The secondary antibody used is HRP goat anti-rabbit IgG H&L (ab97051) at 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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ab180327 (purified) at 1/20 immunoprecipitating CRSP8 in 10 μg PC-12 whole cell lysate (Lanes 1 and 2, observed at 55 kDa). Lane 3 - PBS. For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1500 dilution. Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST
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All lanes : Anti-APG5L/ATG5 antibody [EPR1755(2)] (ab108327) at 1/1000 dilution (unpurified)
Lane 1 : Raji cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : HT-1080 cell lysate
Lane 4 : Human fetal kidney cell lysate
Lane 5 : C6 cell lysate
Lane 6 : Raw264.7 cell lysate
Lane 7 : PC-12 cell lysate
Lane 8 : NIH3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 32 kDaSecondary antibody - anti-rabbit HRP (ab6721)
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Unpurified ab108327, at 1/100 dilution, staining APG5L/ATG5 in paraffin-embedded Human hepatocellular carcinoma tissue by Immunohistochemistry.
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (193)
ab108327 被引用在 193 文献中.
- Ji P et al. Genetically engineered probiotics as catalytic glucose depriver for tumor starvation therapy. Mater Today Bio 18:100515 (2023). PubMed: 36582449
- Livingston MJ et al. Tubular cells produce FGF2 via autophagy after acute kidney injury leading to fibroblast activation and renal fibrosis. Autophagy 19:256-277 (2023). PubMed: 35491858
- Zhang TM et al. TOLLIP-mediated autophagic degradation pathway links the VCP-TMEM63A-DERL1 signaling axis to triple-negative breast cancer progression. Autophagy 19:805-821 (2023). PubMed: 35920704
- Chen H et al. Inhibition of ANGPT2 activates autophagy during hypertrophic scar formation via PI3K/AKT/mTOR pathway. An Bras Dermatol 98:26-35 (2023). PubMed: 36272879
- Schlotawa L et al. An inducible expression system for the manipulation of autophagic flux in vivo. Autophagy 19:1582-1595 (2023). PubMed: 36310368