重组Anti-ATG9A抗体[EPR2450(2)] - BSA and Azide free (ab223528)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR2450(2)] to ATG9A - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IHC-P, IP, 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-ATG9A抗体[EPR2450(2)] - BSA and Azide free
参阅全部 ATG9A 一抗 -
描述
兔单克隆抗体[EPR2450(2)] to ATG9A - BSA and Azide free -
宿主
Rabbit -
特异性
The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
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经测试应用
适用于: Flow Cyt (Intra), IHC-P, IP, WB, ICC/IFmore 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
- HepG2, 293T, A375, cell line lysates Mouse brain and rat brain cell lysates Paraffin-embedded human colon tissue
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常规说明
ab223528 is the carrier-free version of ab108338.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR2450(2) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Anti-ATG9A antibody [EPR2450(2)] (ab108338)
- Alexa Fluor® 488 Anti-ATG9A antibody [EPR2450(2)] (ab206252)
- Alexa Fluor® 647 Anti-ATG9A antibody [EPR2450(2)] (ab206253)
- Alexa Fluor® 555 Anti-ATG9A antibody [EPR2450(2)] (ab211969)
- APC Anti-ATG9A antibody [EPR2450(2)] (ab310808)
- PE Anti-ATG9A antibody [EPR2450(2)] (ab310890)
- Alexa Fluor® 594 Anti-ATG9A antibody [EPR2450(2)] (ab311672)
- Alexa Fluor® 568 Anti-ATG9A antibody [EPR2450(2)] (ab312947)
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab223528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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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IP |
Use at an assay dependent concentration.
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 94 kDa.
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ICC/IF |
Use at an assay dependent concentration.
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说明 |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
IP
Use at an assay dependent concentration. |
WB
Use at an assay dependent concentration. Predicted molecular weight: 94 kDa. |
ICC/IF
Use at an assay dependent concentration. |
靶标
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功能
Involved in autophagy and cytoplasm to vacuole transport (Cvt) vesicle formation. Plays a key role in the organization of the preautophagosomal structure/phagophore assembly site (PAS), the nucleating site for formation of the sequestering vesicle. Cycles between a juxta-nuclear trans-Golgi network compartment and late endosomes. Nutrient starvation induces accumulation on autophagosomes. Starvation-dependent trafficking requires ULK1, ATG13 and SUPT20H. -
序列相似性
Belongs to the ATG9 family. -
细胞定位
Cytoplasmic vesicle, autophagosome membrane. Golgi apparatus, trans-Golgi network membrane. Late endosome membrane. Endoplasmic reticulum membrane. Under amino acid starvation or rapamycin treatment, redistributes from a juxtanuclear clustered pool to a dispersed peripheral cytosolic pool. The starvation-induced redistribution depends on ULK1, ATG13, as well as SH3GLB1. - Information by UniProt
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数据库链接
- Entrez Gene: 79065 Human
- Entrez Gene: 245860 Mouse
- Entrez Gene: 363254 Rat
- Omim: 612204 Human
- SwissProt: Q7Z3C6 Human
- SwissProt: Q68FE2 Mouse
- SwissProt: Q5FWU3 Rat
- Unigene: 323363 Human
see all -
别名
- APG9 autophagy 9-like 1 antibody
- APG9 like 1 antibody
- APG9-like 1 antibody
see all
图片
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).
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This western blot data was generated using the same anti-ATG9A clone (EPR2450(2)] in a different buffer formulation (cat# 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.
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. -
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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).
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Unpurified ab108338 staining ATG9A in 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)
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).
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Unpurified ab108338, at 1/100, staining ATG9A in paraffin-embedded Human colon tissue by Immunohistochemistry.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab108338).
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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Datasheet download
Certificate of Compliance
文献 (7)
ab223528 被引用在 7 文献中.
- Wu X et al. Autophagy regulates Notch degradation and modulates stem cell development and neurogenesis. Nat Commun 7:10533 (2016). ICC/IF ; Human . PubMed: 26837467
- Moreau K et al. Methods to analyze SNARE-dependent vesicular fusion events that regulate autophagosome biogenesis. Methods 75:19-24 (2015). PubMed: 25461811
- Uemura T et al. A cluster of thin tubular structures mediates transformation of the endoplasmic reticulum to autophagic isolation membrane. Mol Cell Biol 34:1695-706 (2014). PubMed: 24591649
- Mitzel DN et al. Age-enhanced endoplasmic reticulum stress contributes to increased Atg9A inhibition of STING-mediated IFN-ß production during Streptococcus pneumoniae infection. J Immunol 192:4273-83 (2014). PubMed: 24670807
- Zavodszky E et al. Mutation in VPS35 associated with Parkinson's disease impairs WASH complex association and inhibits autophagy. Nat Commun 5:3828 (2014). ICC/IF ; Human . PubMed: 24819384
- Moreau K et al. PICALM modulates autophagy activity and tau accumulation. Nat Commun 5:4998 (2014). ICC/IF ; Human . PubMed: 25241929
- Puri C et al. Diverse autophagosome membrane sources coalesce in recycling endosomes. Cell 154:1285-99 (2013). PubMed: 24034251