重组Anti-MTH1抗体[EPR15934-50] (ab200832)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR15934-50] to MTH1
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
概述
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产品名称
Anti-MTH1抗体[EPR15934-50]
参阅全部 MTH1 一抗 -
描述
兔单克隆抗体[EPR15934-50] to MTH1 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IFmore details -
种属反应性
与反应: Human -
免疫原
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: HEK-293T, HAP1, HeLa and Jurkat whole cell lysate; Human fetal kidney and fetal thymus lysates. IHC-P: Human thymus and squamous cell carcinoma of lung tissues. ICC/IF: Jurkat and A549 cells. Flow Cyt (intra): Jurkat cells. IP: Jurkat whole cell lysate.
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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 (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR15934-50 -
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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KO cell lysates
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab200832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) |
1/150.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/5000. Detects a band of approximately 18 kDa (predicted molecular weight: 23 kDa).
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IHC-P |
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/50.
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ICC/IF |
1/500.
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说明 |
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Flow Cyt (Intra)
1/150. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/5000. Detects a band of approximately 18 kDa (predicted molecular weight: 23 kDa). |
IHC-P
1/50. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/50. |
ICC/IF
1/500. |
靶标
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功能
Antimutagenic. Acts as a sanitizing enzyme for oxidized nucleotide pools, thus suppressing cell dysfunction and death induced by oxidative stress. Hydrolyzes 8-oxo-dGTP, 8-oxo-dATP and 2-OH-dATP, thus preventing misincorporation of oxidized purine nucleoside triphosphates into DNA and subsequently preventing A:T to C:G and G:C to T:A transversions. Able to hydrolyze also the corresponding ribonucleotides, 2-OH-ATP, 8-oxo-GTP and 8-oxo-ATP. Does not play a role in U8 snoRNA decapping activity. Binds U8 snoRNA. -
组织特异性
Widely expressed with highest expression in thymus, testis, embryo and proliferating blood lymphocytes. -
序列相似性
Belongs to the Nudix hydrolase family.
Contains 1 nudix hydrolase domain. -
发展阶段
In peripheral blood lymphocytes, expressed at much higher levels in proliferating cells than in resting cells. -
翻译后修饰
The N-terminus is blocked. -
细胞定位
Cytoplasm. Mitochondrion matrix and Cytoplasm. Mitochondrion matrix. Nucleus. Mostly present in cytoplasm. Variant Met-124 has decreased efficiency in translocation to mitochondria. - Information by UniProt
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数据库链接
- Entrez Gene: 4521 Human
- Omim: 600312 Human
- SwissProt: P36639 Human
- Unigene: 534331 Human
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别名
- 2-hydroxy-dATP diphosphatase antibody
- 7 8 dihydro 8 oxoguanine triphosphatase antibody
- 7 antibody
see all
图片
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All lanes : Anti-MTH1 antibody [EPR15934-50] (ab200832) at 1/5000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : NUDT1 knockout HEK-293T cell lysate
Lane 3 : HAP1 cell lysate
Lane 4 : HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 23 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?Lanes 1- 4: Merged signal (red and green). Green - ab200832 observed at 18 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab200832 was shown to react with MTH1 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266400 (knockout cell lysate ab257565) was used. Wild-type HEK-293T and NUDT1 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab200832 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 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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Immunocytochemistry/Immunofluorescence analysis of Daudi (human Burkitt's lymphoma) labelling MTH-1 with purified ab200832 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (Ab150077). Nuclei counterstained with DAPI (blue).
Control: PBS only
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: MTH1 knockout HAP1 cell lysate (20 µg)
Lane 3: Jurkat cell lysate (20 µg)
Lane 4: A549 cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab200832 observed at 18 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab200832 was shown to specifically react with MTH1 when MTH1 knockout samples were used. Wild-type and MTH1 knockout samples were subjected to SDS-PAGE. ab200832 at a dilution of 1/5000 and ab8245 (loading control to GAPDH) diluted to 1/10,000 were 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 hour at room temperature before imaging. -
Anti-MTH1 antibody [EPR15934-50] (ab200832) at 1/5000 dilution + Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate at 20 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 23 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?
Exposure time: 15 secondsBlocking/dilution Buffer: 5% NFDM/TBST.
The expression profile observed is consistent with the following literature PMID: 11296483.
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All lanes : Anti-MTH1 antibody [EPR15934-50] (ab200832) at 1/5000 dilution
Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal thymus lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 23 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteBlocking/dilution Buffer: 5% NFDM/TBST.
The expression profile observed is consistent with the following literature PMID: 11296483.
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Immunohistochemical analysis of paraffin-embedded Human thymus tissue labeling MTH1 with ab200832 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on Human thymus tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemical analysis of paraffin-embedded Human squamous cell carcinoma of lung tissue labeling MTH1 with ab200832 at 1/100 dilution followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on Human squamous cell carcinoma of lung tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
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 2% paraformaldehyde-fixed Jurkat (Human T cell leukemia cells from peripheral blood)cells labeling MTH1 with ab200832 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/150 dilution was used as the secondary antibody.
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MTH1 was immunoprecipitated from 1mg of Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysate with ab200832 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab200832 at 1/2000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.
Lane 1: Jurkat whole cell lysate 10ug (Input). Lane 2: ab200832 IP in Jurkat whole cell lysate. Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab200832 in Jurkat whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.Exposure time: 3 seconds.
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 (4)
ab200832 被引用在 4 文献中.
- Chen Y et al. MutT Homolog 1 Inhibitor Karonudib Attenuates Autoimmune Hepatitis by Inhibiting DNA Repair in Activated T Cells. Hepatol Commun 6:1016-1031 (2022). PubMed: 34894107
- Ou Q et al. Nudix hydrolase 1 is a prognostic biomarker in hepatocellular carcinoma. Aging (Albany NY) 12:7363-7379 (2020). PubMed: 32341205
- Qing X et al. Anticancer effect of (S)-crizotinib on osteosarcoma cells by targeting MTH1 and activating reactive oxygen species. Anticancer Drugs 29:341-352 (2018). PubMed: 29420337
- Ahmed W & Lingner J PRDX1 and MTH1 cooperate to prevent ROS-mediated inhibition of telomerase. Genes Dev 32:658-669 (2018). PubMed: 29773556