Anti-Topoisomerase I抗体(ab3825)
Key features and details
- Rabbit polyclonal to Topoisomerase I
- Suitable for: WB, ICC
- Reacts with: Human
- Isotype: IgG
概述
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产品名称
Anti-Topoisomerase I抗体
参阅全部 Topoisomerase I 一抗 -
描述
兔多克隆抗体to Topoisomerase I -
宿主
Rabbit -
经测试应用
适用于: WB, ICCmore details -
种属反应性
与反应: Human -
免疫原
Recombinant fragment corresponding to Human Topoisomerase I aa 401-600.
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常规说明
This product can be used as part of an assay for sumoylation activity. Human Aos 1 + Uba 2 (ab3804), Ubc 9 (ab3803) and Sumo 1 (ab3801) can be used to promote in vitro sumoylation of a sumoylation marker (human Topoisomerase I protein fragment) (ab3828). The reaction products can be detected using our Sumo 1 (ab3819 and ab3824) and Topoisomerase I (ab3825) antibodies. Sumoylation assays are carried out in a final volume of 20µl in reaction conditions (20 mM Hepes pH 7.5, 5mM MgCl2, 2mM ATP). This antibody is equivalent to the antibody used in our Sumoylation Control kits. Sumoylation Protocol: 1. Prepare a suitable purified substrate protein. (For the control, use 2µl Topoisomerase I marker for each reaction.) 2. In each reaction, add 4µl E2 to substrate first, then 2µl Sumo 1, 2µl 10x reaction buffer, 2µl E1. Finally, add H2O to bring up to 20µl. We would recommend adding fresh 2mM ATP to be sure that sufficient energy is supplied. 3. The best reaction concentration of proteins is as following: Aos 1 + Uba 2: 7.5µg/ml. Ubc 9: 50µg/ml. SUMO 1: 50µg/ml. For the control assay we recommend running the assay at 37ºC for 30-60 minutes. 4. Detect the reaction products by Western blot using a suitable antibody. For the control reaction use 1/1000 dilution of the supplied Topoisomerase I antibody. Four sumoylated bands should be seen on the gel for the control reaction. This assay has been shown to work with crude extracts. Be aware that Uba 2 contains his-rich regions which might cross-react with antibodies against the 6x-His epitope tag. During western analysis with anti-6x-His antibodies, Uba 2 at 80 kDa might be shown.
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
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 or -80°C. Avoid freeze / thaw cycle. -
存储溶液
Constituent: 100% PBS -
Concentration information loading... -
纯度
Whole antiserum -
Primary antibody说明
This product can be used as part of an assay for sumoylation activity. Human Aos 1 + Uba 2 (ab3804), Ubc 9 (ab3803) and Sumo 1 (ab3801) can be used to promote in vitro sumoylation of a sumoylation marker (human Topoisomerase I protein fragment) (ab3828). The reaction products can be detected using our Sumo 1 (ab3819 and ab3824) and Topoisomerase I (ab3825) antibodies. Sumoylation assays are carried out in a final volume of 20µl in reaction conditions (20 mM Hepes pH 7.5, 5mM MgCl2, 2mM ATP). This antibody is equivalent to the antibody used in our Sumoylation Control kits. Sumoylation Protocol: 1. Prepare a suitable purified substrate protein. (For the control, use 2µl Topoisomerase I marker for each reaction.) 2. In each reaction, add 4µl E2 to substrate first, then 2µl Sumo 1, 2µl 10x reaction buffer, 2µl E1. Finally, add H2O to bring up to 20µl. We would recommend adding fresh 2mM ATP to be sure that sufficient energy is supplied. 3. The best reaction concentration of proteins is as following: Aos 1 + Uba 2: 7.5µg/ml. Ubc 9: 50µg/ml. SUMO 1: 50µg/ml. For the control assay we recommend running the assay at 37ºC for 30-60 minutes. 4. Detect the reaction products by Western blot using a suitable antibody. For the control reaction use 1/1000 dilution of the supplied Topoisomerase I antibody. Four sumoylated bands should be seen on the gel for the control reaction. This assay has been shown to work with crude extracts. Be aware that Uba 2 contains his-rich regions which might cross-react with antibodies against the 6x-His epitope tag. During western analysis with anti-6x-His antibodies, Uba 2 at 80 kDa might be shown. -
克隆
多克隆 -
同种型
IgG -
研究领域
相关产品
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ChIP Related Products
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Compatible Secondaries
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Isotype control
应用
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (9) |
1/1000. Predicted molecular weight: 91 kDa.
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| ICC |
1/50 - 1/100.
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| 说明 |
|---|
|
WB
1/1000. Predicted molecular weight: 91 kDa. |
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ICC
1/50 - 1/100. |
靶标
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功能
The reaction catalyzed by topoisomerases leads to the conversion of one topological isomer of DNA to another. -
疾病相关
Note=A chromosomal aberration involving TOP1 is found in a form of therapy-related myelodysplastic syndrome. Translocation t(11;20)(p15;q11) with NUP98. -
序列相似性
Belongs to the eukaryotic type I topoisomerase family. -
翻译后修饰
Sumoylated. Lys-117 is the main site of sumoylation. Sumoylation plays a role in partitioning TOP1 between nucleoli and nucleoplasm. Levels are dramatically increased on camptothecin (CPT) treatment. -
细胞定位
Nucleus > nucleolus. Nucleus > nucleoplasm. Diffuse nuclear localization with some enrichment in nucleoli. On CPT treatment, cleared from nucleoli into nucleoplasm. Sumolyated forms found in both nucleoplasm and nucleoli. - Information by UniProt
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数据库链接
- Entrez Gene: 7150 Human
- Omim: 126420 Human
- SwissProt: P11387 Human
- Unigene: 472737 Human
- Unigene: 712686 Human
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别名
- DNA topoisomerase 1 antibody
- DNA topoisomerase I antibody
- NUP98 fusion gene antibody
see all
图片
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Immunocytochemistry - Anti-Topoisomerase I antibody (ab3825)This image is courtesy of an anonymous Abreview
ab3825 staining Topoisomerase I in human 293T cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 3% BSA for 1 hour at 22°C. Samples were incubated with primary antibody (1/100 in PBS + 1% BSA) for 1 hour at 22°C. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG polyclonal (1/1000) was used as the secondary antibody.
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Immunocytochemistry - Anti-Topoisomerase I antibody (ab3825)blue: DAPI; green: Topo I. Hela cell staining. -
Western blot - Anti-Topoisomerase I antibody (ab3825)Primary antibody incubated at a 1/1000 dilution for 60 minutes at 25ºC.
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Western blot - Anti-Topoisomerase I antibody (ab3825)All lanes : Anti-Topoisomerase I antibody (ab3825) at 1/5000 dilution
Lane 1 : SW210.5 human SCLC cell xenograft lysate.
Lane 2 : 5M2 human NSCLC cell lysate (bicarb. buffer).
Lane 3 : 5M2 human NSCLC cell lysate (M-Per buffer).
Lane 4 : Normal human lung tissue lysate.
Lane 5 : A431 cell (human epidermoid carcinoma) lysate .
Lane 6 : Rh30 cell (human rhabdomyosarcoma) lysate.
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti rabbit heavy and light (HRP) at 1/25000 dilution
Performed under reducing conditions.
Predicted band size: 91 kDa
This image is courtesy of an Abreview submitted by Mike Campa on 7 September 2005. We do not have any further information relating to this image. -
Western blot - Anti-Topoisomerase I antibody (ab3825)This image is courtesy of an anonymous AbreviewAll lanes : Anti-Topoisomerase I antibody (ab3825) at 1/200 dilution
Lane 1 : K562 nuclear lysate
Lane 2 : Mouse N2a whole cell lysate at 100 µg
Secondary
All lanes : Donkey anti-rabbit IgG HRP polyclonal at 1/1000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 91 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Additional bands at: 35 kDa (possible non-specific binding)
Exposure time: 15 seconds
实验方案
数据表及文件
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SDS download
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Datasheet download
文献 (22)
ab3825 被引用在 22 文献中.
- Sheraz M et al. Cellular DNA Topoisomerases Are Required for the Synthesis of Hepatitis B Virus Covalently Closed Circular DNA. J Virol 93:N/A (2019). PubMed: 30867306
- Hazan I et al. Activation of Oncogenic Super-Enhancers Is Coupled with DNA Repair by RAD51. Cell Rep 29:560-572.e4 (2019). PubMed: 31618627
- Ahmad M et al. Topoisomerase 3ß is the major topoisomerase for mRNAs and linked to neurodevelopment and mental dysfunction. Nucleic Acids Res 45:2704-2713 (2017). PubMed: 28039324
- Bansal K et al. The transcriptional regulator Aire binds to and activates super-enhancers. Nat Immunol 18:263-273 (2017). PubMed: 28135252
- Wan X et al. SIRT1-PGC1a-NF?B Pathway of Oxidative and Inflammatory Stress during Trypanosoma cruzi Infection: Benefits of SIRT1-Targeted Therapy in Improving Heart Function in Chagas Disease. PLoS Pathog 12:e1005954 (2016). WB ; Mouse . PubMed: 27764247