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  4. TOP1 Recombinant Monoclonal Antibody

TOP1 Recombinant Monoclonal Antibody

  • 货号：
    CSB-RA792129A0HU
  • 规格：
    ￥1320
  • 图片：
    • Western Blot
      Positive WB detected in: Hela whole cell lysate, MCF-7 whole cell lysate, K562 whole cell lysate, HL60 whole cell lysate, PC-3 whole cell lysate
      All lanes: TOP1 antibody at 1:2000
      Secondary
      Goat polyclonal to rabbit IgG at 1/50000 dilution
      Predicted band size: 91 kDa
      Observed band size: 91 kDa
    • IHC image of CSB-RA792129A0HU diluted at 1:100 and staining in paraffin-embedded human small intestine tissue performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.
    • IHC image of CSB-RA792129A0HU diluted at 1:100 and staining in paraffin-embedded human colon cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a Goat anti-rabbit IgG polymer labeled by HRP and visualized using 0.05% DAB.
    • Overlay histogram showing HepG2 cells stained with CSB-RA792129A0HU (red line) at 1:50. The cells were fixed with 70% Ethylalcohol (18h) and then incubated in 10% normal goat serum to block non-specific protein-protein interactions followedby the antibody (1µg/1*106 cells) for 1 h at 4℃.The secondary antibody used was FITC-conjugated goat anti-rabbit IgG (H+L) at 1/200 dilution for 30min at 4℃. Control antibody (green line) was Rabbit IgG (1µg/1*106 cells) used under the same conditions. Acquisition of >10,000 events was performed.
    • Immunoprecipitating TOP1 in K562 whole cell lysate
      Lane 1: Rabbit control IgG instead of CSB-RA792129A0HU in K562 whole cell lysate. For western blotting,a HRP-conjugated Protein G antibody was used as the secondary antibody (1/2000)
      Lane 2: CSB-RA792129A0HU(2µg)+ K562 whole cell lysate(500µg)
      Lane 3: K562 whole cell lysate (10µg)
  • 其他：

产品详情

  • 产品描述：

    The production of the recombinant TOP1 antibody depended on Single B Cell technology. There are 3 main steps in the production: 1, Isolation of single B cells. High-throughput methods could be used to obtain the efficient identification and desired specificity of B cells. 11, Single B cell antibody sequencing and cloning. In this step, the antibody gene sequence of TOP1 was obtained and introduced to plasmids, which then would be transferred to mammalian cells for in vitro expression of the TOP1 antibody. 3, Screening of antibodies. The target antibody was obtained in this step. And it has been validated in ELISA, WB, IHC, FC, IP.

    TOP1, also known as swivelase or untwisting enzyme, relaxes both positive and negative supercoils by nicking DNA, allowing the broken DNA strand to rotate around the complementary intact strand of the DNA duplex before catalyzing the nick closure. This action is especially crucial during transcription because helical restrictions might obstruct DNA replication and transcription, halting cell growth. TOP1 plays an essential role in normal development in mammals. TOP1 and TOP2A are tumor drivers in a variety of cancers, making them interesting and effective targets for anti-cancer drug development.

  • Uniprot No.：
    P11387
  • 基因名：
  • 别名：
    DNA topoisomerase 1 (EC 5.99.1.2) (DNA topoisomerase I), TOP1
  • 反应种属：
    Human
  • 免疫原：
    A synthesized peptide derived from human TOP1
  • 免疫原种属：
    Homo sapiens (Human)
  • 标记方式：
    Non-conjugated
  • 克隆类型：
    Monoclonal
  • 抗体亚型：
    Rabbit IgG
  • 纯化方式：
    Affinity-chromatography
  • 克隆号：
    6D8
  • 浓度：
    It differs from different batches. Please contact us to confirm it.
  • 保存缓冲液：
    Rabbit IgG in phosphate buffered saline, pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol.
  • 产品提供形式：
    Liquid
  • 应用范围：
    ELISA, WB, IHC, FC, IP
  • 推荐稀释比：
    Application Recommended Dilution
    WB 1:500-1:5000
    IHC 1:50-1:200
    FC 1:20-1:200
    IP 1:200-1:1000
  • Protocols：
  • 储存条件：
    Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
  • 货期：
    Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.

产品评价

靶点详情

  • 功能：
    Releases the supercoiling and torsional tension of DNA introduced during the DNA replication and transcription by transiently cleaving and rejoining one strand of the DNA duplex. Introduces a single-strand break via transesterification at a target site in duplex DNA. The scissile phosphodiester is attacked by the catalytic tyrosine of the enzyme, resulting in the formation of a DNA-(3'-phosphotyrosyl)-enzyme intermediate and the expulsion of a 5'-OH DNA strand. The free DNA strand then rotates around the intact phosphodiester bond on the opposing strand, thus removing DNA supercoils. Finally, in the religation step, the DNA 5'-OH attacks the covalent intermediate to expel the active-site tyrosine and restore the DNA phosphodiester backbone. Regulates the alternative splicing of tissue factor (F3) pre-mRNA in endothelial cells. Involved in the circadian transcription of the core circadian clock component ARNTL/BMAL1 by altering the chromatin structure around the ROR response elements (ROREs) on the ARNTL/BMAL1 promoter.
  • 基因功能参考文献：
    1. TOP1 differentially modulates R-loops across the human genome. PMID: 30060749
    2. Results from a study on gene expression variability markers in early-stage human embryos shows that TOP1 is a putative expression variability marker for the 3-day, 8-cell embryo stage. PMID: 26288249
    3. High TOP1 expression is associated with aggressive tumor phenotype in breast cancer. PMID: 26959889
    4. Ru/Fe bimetallic complexes: Synthesis, characterization, cytotoxicity and study of their interactions with DNA/HSA and human topoisomerase IB. PMID: 29107586
    5. two regions of topoisomerase I, the N-terminal and the linker domains, were critical for subnuclear localization of the enzyme. The linker domain and the distal region of the N-terminal domain directed topoisomerase I to the nucleolus, whereas the remaining region of the N-terminal domain was responsible for the nucleoplasmic localization. PMID: 27428351
    6. High TOP1 expression is associated with Colorectal Cancers. PMID: 28870917
    7. HMGA2 potentiates the clinically important topoisomerase I inhibitor irinotecan/SN-38 in trapping the enzyme in covalent DNA-complexes, thereby attenuating transcription. PMID: 27587582
    8. IMMP2L transcription requires Topoisomerase I in human primary astrocytes PMID: 27932244
    9. This work identifies ADP-ribose polymers as key determinant for regulating Top1 subnuclear dynamics. PMID: 27466387
    10. copper(II) thiosemicarbazone complex may hit human topoisomerase IB and that metal coordination can be useful to improve cytotoxicity of this versatile class of compounds PMID: 27431056
    11. High prevalence of anti-topoisomerase I antibody positivity was found among Thai systemic sclerosis patients and this was associated with a high frequency of hand deformity, ACA negativity, a short duration of pulmonary fibrosis in diffuse cutaneous and a lower frequency of Raynaud's phenomenon in limited cutaneous subsets. PMID: 25293362
    12. These findings reveal BAZ1B as a key facilitator of topoisomerase I function during DNA replication that affects the response of cancer cells to topoisomerase I inhibitors. PMID: 27050524
    13. Results identified TOP1 gene copy gain, a loss of chromosome 20, and new yet unreported TOP1 mutations (R364K and G717R) in close proximity to the SN-38 binding site conferring colon cancer cells resistance to the drug. PMID: 27029323
    14. the highest expression of GGH and EGFR was noted in the left-sided colon; the highest expression of DHFR, FPGS, TOP1 and ERCC1 was noted in the rectosigmoid, whereas TYMP expression was approximately equivalent in the right-sided colon and rectum PMID: 26676887
    15. We found that the resistant cell lines showed 7-100 fold increased resistance to SN-38 but remained sensitive to docetaxel and the non-camptothecin Top1 inhibitor LMP400 PMID: 26801902
    16. Data show that the single-molecule supercoil relaxation assay is a sensitive method to elucidate the detailed mechanisms of type IB topoisomerase (Top1) inhibitors and is relevant for the cellular efficacy of Top1 inhibitors. PMID: 26351326
    17. TOP1 bound at promoters was discovered to become fully active only after pause-release. This transition coupled the phosphorylation of the carboxyl-terminal-domain (CTD) of RNA polymerase II (RNAPII) with stimulation of TOP1 above its basal rate, enhancing its processivity. PMID: 27058666
    18. Data show that inhibition of DNA-dependent protein kinase catalytic subunit (DNA-PK) prevents type I DNA topoisomerase (Top1) degradation and proteasome activity in camptothecin (CPT)-treated quiescent WI38 cells. PMID: 26578593
    19. NO-induced down-regulation of topoisomerase I protein. PMID: 26540186
    20. b-ELE could inhibit the proliferation of HepG-2 cells and interfere with the expression and activity of TOPO I and TOPO IIa PMID: 26221582
    21. Acridine derivatives inhibited DNA topoisomerases I and II and prevented proliferation of HL-60 cells. PMID: 25960253
    22. Topoisomerase-1 and -2A gene copy numbers are elevated in mismatch repair-proficient colorectal cancers. PMID: 25777966
    23. Cinobufacini inhibited the proliferation of the HepG-2 cells induced apoptosis in a dose- and time-dependent manner and downregulated the mRNA and protein expression levels of TOPO I and TOPO II PMID: 25815590
    24. YB-1 serves as an intracellular promoter of TOPO1 catalytic activity that enhances camptothecin sensitivity through its direct interaction with TOPO1. PMID: 25539742
    25. We here report that a substantial number of patients with bile duct or pancreatic cancer have increased TOP1 copy number and increased TOP1/centromere-20 ratio. PMID: 25615400
    26. Results show that TOP1 gene is amplified in a subset of breast cancer patients suggesting the gene as a potential biomarker for response to treatment with Top1 inhibitors. PMID: 25855483
    27. Results show that camptothecin resistance status of colorectal cancer cells depend on the phosphorylation of TOP1 and the presence of CD44. PMID: 24960044
    28. varying expression levels of TOP1 and TDP1 polypeptides in multiple colorectal cancer cell lines and in clinical colorectal cancer samples, are reported. PMID: 25522766
    29. CCR6 SNPs are a risk factor for presence of anti-topoisomerase I antibodies in systemic sclerosis. PMID: 26314374
    30. ERCC1-XPF participates in DNA repair of the Top1-DNA damage complex. PMID: 26025908
    31. both TOP1 and TDP1 were upregulated in the tumor tissue compared to the adjacent non-tumor tissue in non-small cell lung cancer tissue PMID: 25987486
    32. mutations at the hydrophobic or charged residues of the putative polymer binding sites do not interfere with the ability of poly(ADP-ribose) to antagonize the antitumor activity of topoisomerase I poisons. PMID: 25227992
    33. No TOP1 copy number changes were found in patients with de novo diffuse large B-cell lymphoma or relapsed DLBCL treated with chemotherapy regimens including TOP2-targeting drugs. PMID: 25931012
    34. Mutation of Gly717Phe in human topoisomerase 1B has an effect on enzymatic function, reactivity to the camptothecin anticancer drug and on the linker domain orientation PMID: 25910424
    35. Top1 cleavage events generate short deletions; Top1 also promotes nick religation at rNMP sites PMID: 25887397
    36. analysis of how human and yeast DNA topoisomerase I domain interactions impact enzyme activity and sensitivity to camptothecin PMID: 25795777
    37. Study describes a molecular mechanism that operates at functional androgen-regulated enhancers and identify DNA topoisomerase I as a critical DNA-nicking enzyme involved in the process of cell-specific, ligand-driven enhancer activation. PMID: 25619691
    38. The combination of Top1 inhibitors with VE-821 inhibited the phosphorylation of ATR and Chk1. PMID: 25269479
    39. DNA topoisomerases I and II are involved in initiating APH-induced common fragile site breakage at RET PMID: 24040417
    40. Collectively, these findings suggest that the combinatorial inhibition of MET and Top1 is a potentially efficacious treatment strategy for Small cell lung cancer . PMID: 24327519
    41. our data show for the first time that HIF-1alpha is strongly correlated with resistance to topoisomerase I inhibitors in hepatocellular carcinoma. PMID: 24362462
    42. Analysis of the dynamical properties of DNA topoisomerase 1B bound to the more biologically relevant supercoiled substrate reveals an increased number of protein-DNA interactions and, most strikingly, the presence of a secondary protein-DNA binding site. PMID: 25056319
    43. homologous recombination seems relevant especially for Top1 and, to a lesser extent, for Top2 inhibitors. We also found and discuss differential pathways among Top1 inhibitors and Top2 inhibitors PMID: 24130054
    44. key factors in a molecular pathway connecting Top1 inhibition and human HIF-1alpha protein regulation and activity, widening the biologic and molecular activity of camptothecin PMID: 24252850
    45. [review] Telangiectasia and anti-CENP or anti-topo I antibodies in the presence of Raynaud's phenomenon and proximal skin thickening increase the sensitivity of a very early diagnosis of systemic sclerosis from 57% to 97%. PMID: 24461384
    46. This property facilitates the deprotonation of the 5' DNA end, suggesting that this is the limiting step in the topoisomerase religation mechanism. PMID: 23368812
    47. These results suggest that topo I is a novel target of erlotinib and a combination of TKIs with topo I inhibitors may be an effective treatment for breast cancer. PMID: 24399039
    48. Critical endogenous pathogenic lesions are associated with neurodegenerative syndromes arising from aberrant TOP-1 DNA. PMID: 24793032
    49. A fully functional linker is required to confer camptothecin sensitivity to topoisomerase I. PMID: 24004603
    50. MiR-23a could directly bind to 3'untranslated region of TOP1 mRNA, and suppress the corresponding protein expression. PMID: 24103454

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  • 相关疾病：
    A chromosomal aberration involving TOP1 is found in a form of therapy-related myelodysplastic syndrome. Translocation t(11;20)(p15;q11) with NUP98.
  • 亚细胞定位：
    Nucleus, nucleolus. Nucleus, nucleoplasm. Note=Diffuse nuclear localization with some enrichment in nucleoli. On CPT treatment, cleared from nucleoli into nucleoplasm. Sumoylated forms found in both nucleoplasm and nucleoli.
  • 蛋白家族：
    Type IB topoisomerase family
  • 组织特异性：
    Endothelial cells.
  • 数据库链接：

    HGNC: 11986

    OMIM: 126420

    KEGG: hsa:7150

    STRING: 9606.ENSP00000354522

    UniGene: Hs.472737