重组Anti-Indoleamine 2,3-dioxygenase抗体[EPR20374] (ab211017)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20374] to Indoleamine 2, 3-dioxygenase
- Suitable for: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-P
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-Indoleamine 2,3-dioxygenase抗体[EPR20374]
参阅全部 Indoleamine 2, 3-dioxygenase 一抗 -
描述
兔单克隆抗体[EPR20374] to Indoleamine 2,3-dioxygenase -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, ICC/IF, IP, IHC-Pmore details -
种属反应性
与反应: Human -
免疫原
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: Wild-type A549 Treated IFN gamma, Human ovary cancer, placenta and tonsil lysates; SK-OV-3 whole cell lysate; HeLa whole cell lysate treated with 50ng/ml Interferon-gamma (IFN-gamma) for 16 hours. IHC-P: Human spleen, tonsil, placenta and endometrium cancer tissues. ICC/IF: HeLa cells treated with IFN-gamma (50 ng/ml) for 16 hours. Flow Cyt (intra): HeLa cells treated with IFN-gamma (50 ng/ml) for 16 hours. IP: HeLa whole cell lysate treated with 50ng/ml IFN-gamma for 16h.
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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 long term. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR20374 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
- Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - Low endotoxin, Azide free (ab224263)
- Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] - BSA and Azide free (ab271990)
- Alexa Fluor® 647 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab281580)
- Alexa Fluor® 555 Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab281769)
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Compatible Secondaries
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Conjugation kits
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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ab211017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) |
1/500.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (1) |
1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa).
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ICC/IF |
1/2000.
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IP |
1/40.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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说明 |
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Flow Cyt (Intra)
1/500. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000. Detects a band of approximately 45 kDa (predicted molecular weight: 45 kDa). |
ICC/IF
1/2000. |
IP
1/40. |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
靶标
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功能
Catalyzes the cleavage of the pyrrol ring of tryptophan and incorporates both atoms of a molecule of oxygen. -
通路
Amino-acid degradation; L-tryptophan degradation via kynurenine pathway; L-kynurenine from L-tryptophan: step 1/2. -
序列相似性
Belongs to the indoleamine 2,3-dioxygenase family. - Information by UniProt
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数据库链接
- Entrez Gene: 3620 Human
- Omim: 147435 Human
- SwissProt: P14902 Human
- Unigene: 840 Human
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别名
- 3-dioxygenase antibody
- I23O1_HUMAN antibody
- IDO 1 antibody
see all
图片
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All lanes : Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017) at 1/1000 dilution
Lane 1 : Wild-type A549 Vehicle Control IFN gamma (0 ng/ml, 48 h) cell lysate
Lane 2 : Wild-type A549 Treated IFN gamma (25 ng/ml, 48 h) cell lysate
Lane 3 : IDO1 knockout A549 Vehicle Control IFN gamma (0 ng/ml, 48 h) cell lysate
Lane 4 : IDO1 knockout A549 Treated IFN gamma (25 ng/ml, 48 h) cell lysate
Lane 5 : SK-OV-3 cell lysate
Lane 6 : MCF7 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 45 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?Lanes 1 - 6: Merged signal (red and green). Green - ab211017 observed at 40 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.
ab211017 was shown to react with Indoleamine 2, 3-dioxygenase in treated wild-type A549 cells in Western blot with no signal observed in treated IDO1 knockout cell line ab266949 (IDO1 knockout cell lysate ab256948). Wild-type A549 and IDO1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab211017 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4 °C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)
Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic and nuclear staining on dendritic cells of human spleen is observed (PMID: 21328335, PMID: 25271151).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017) at 1/1000 dilution
Lane 1 : Human ovary cancer lysate at 20 µg
Lane 2 : Human placenta lysate at 20 µg
Lane 3 : Human tonsil lysate at 20 µg
Lane 4 : SK-OV-3 (Human ovarian cancer cell line) whole cell lysate at 10 µg
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 45 kDa
Observed band size: 45 kDaBlocking/Dilution buffer: 5% NFDM/TBST.
Exposure time: Lane 1-3: 3 minutes; Lane 4: 15 seconds.
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Immunocytochemistry/ Immunofluorescence - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 50ng/ml IFN-γ for 16 hours or untreated, labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
The signal increased after treatment with IFN-γ (50 ng/ml) for 16 hours on HeLa cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594)) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.
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All lanes : Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017) at 1/1000 dilution
Lane 1 : Untreated HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 2 : HeLa whole cell lysate treated with 50ng/ml Interferon-gamma (IFN-gamma, ab51240) for 16 hours
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 45 kDa
Observed band size: 45 kDa
Exposure time: 15 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The level of Indoleamine 2, 3-dioxygenase expression can be induced by IFN-γ treatment (PMID 16368976).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic and nuclear staining on dendritic cells of human tonsil is observed (PMID: 21328335, PMID: 25271151).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)
Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic and nuclear staining on endothelial cells of human placenta is observed (PMID: 21328335, PMID: 25271151).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)
Immunohistochemical analysis of paraffin-embedded human endometrium cancer tissue labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Cytoplasmic and nuclear staining on human endometrium cancer is observed (PMID: 26155395).
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells, treated with 50ng/ml IFN-? for 16h (red) or untreated (green), labeling Indoleamine 2, 3-dioxygenase with ab211017 at 1/500 dilution compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
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Indoleamine 2, 3-dioxygenase was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate treated with 50ng/ml IFN-γ for 16h with ab211017 at 1/40 dilution.
Western blot was performed from the immunoprecipitate using ab211017 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.
Lane 1: HeLa treated with 50ng/ml IFN-γ for 16h whole cell lysate 10 µg (Input).
Lane 2: ab211017 IP in HeLa treated with 50ng/ml IFN-γ for 16h whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab211017 in HeLa treated with 50ng/ml IFN-γ for 16h whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 1 second.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374] (ab211017)
Tissue Microarrays stained for "Anti-Indoleamine 2, 3-dioxygenase antibody [EPR20374]” using "ab211017" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). The sections were incubated with ab211017 at +4°C overnight followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP polymer).
实验方案
数据表及文件
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SDS download
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Datasheet download
Certificate of Compliance
文献 (12)
ab211017 被引用在 12 文献中.
- Chen Y et al. SNF5, a core subunit of SWI/SNF complex, regulates melanoma cancer cell growth, metastasis, and immune escape in response to matrix stiffness. Transl Oncol 17:101335 (2022). PubMed: 34999540
- Qiu T et al. Astragaloside IV Inhibits the Proliferation of Human Uterine Leiomyomas by Targeting IDO1. Cancers (Basel) 14:N/A (2022). PubMed: 36139584
- Shen R et al. Decoding the colorectal cancer ecosystem emphasizes the cooperative role of cancer cells, TAMs and CAFsin tumor progression. J Transl Med 20:462 (2022). PubMed: 36209225
- Li H et al. Indoleamine 2,3-dioxygenase mediates the therapeutic effects of adipose-derived stromal/stem cells in experimental periodontitis by modulating macrophages through the kynurenine-AhR-NRF2 pathway. Mol Metab 66:101617 (2022). PubMed: 36270612
- Someya S et al. Tryptophan Metabolism Regulates Proliferative Capacity of Human Pluripotent Stem Cells. iScience 24:102090 (2021). PubMed: 33615198