使用敲除细胞株进行验证

Anti-Werner's syndrome helicase WRN抗体[8H3] (ab66601)

概述

  • 产品名称Anti-Werner's syndrome helicase WRN抗体[8H3]
    参阅全部 Werner's syndrome helicase WRN 一抗
  • 描述
    小鼠单克隆抗体[8H3] to Werner's syndrome helicase WRN
  • 经测试应用适用于: ICC/IF, IP, WBmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Human WRN helicase containing N-terminal hexahistidine produced in inscet Sf21 cells by baculovirus expression systems as described by Suzuki et al, and purified by affinity column chromatography using Ni-chelating NTA agarose gel.

  • 表位The epitope of 8H3 resides in the N-terminal 232-368 region.

性能

  • 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Preservative: 0.05% Sodium Azide
    Constituents: PBS
  • Concentration information loading...
  • 纯度Protein A purified
  • 克隆单克隆
  • 克隆编号8H3
  • 同种型IgG1
  • 轻链类型kappa
  • 研究领域

相关产品

应用

Our Abpromise guarantee covers the use of ab66601 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF Use a concentration of 1 µg/ml.
IP Use at an assay dependent dilution.
WB 1/500 - 1/1000. Detects a band of approximately 180 kDa (predicted molecular weight: 160 kDa).

靶标

  • 功能Multifunctional enzyme that has both magnesium and ATP-dependent DNA-helicase activity and 3'->5' exonuclease activity towards double-stranded DNA with a 5'-overhang. Has no nuclease activity towards single-stranded DNA or blunt-ended double-stranded DNA. Binds preferentially to DNA substrates containing alternate secondary structures, such as replication forks and Holliday junctions. May play an important role in the dissociation of joint DNA molecules that can arise as products of homologous recombination, at stalled replication forks or during DNA repair. Alleviates stalling of DNA polymerases at the site of DNA lesions. Important for genomic integrity. Plays a role in the formation of DNA replication focal centers; stably associates with foci elements generating binding sites for RP-A.
  • 疾病相关Defects in WRN are a cause of Werner syndrome (WRN) [MIM:277700]. WRN is a rare autosomal recessive progeroid syndrome characterized by the premature onset of multiple age-related disorders, including atherosclerosis, cancer, non-insulin-dependent diabetes mellitus, ocular cataracts and osteoporosis. The major cause of death, at a median age of 47, is myocardial infarction. Currently all known WS mutations produces prematurely terminated proteins.
    Defects in WRN may be a cause of colorectal cancer (CRC) [MIM:114500].
  • 序列相似性Belongs to the helicase family. RecQ subfamily.
    Contains 1 3'-5' exonuclease domain.
    Contains 1 helicase ATP-binding domain.
    Contains 1 helicase C-terminal domain.
    Contains 1 HRDC domain.
  • 翻译后修饰Phosphorylated by PRKDC. Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 细胞定位Nucleus > nucleolus. Nucleus.
  • Information by UniProt
  • 数据库链接
  • 别名
    • DKFZp686C2056 antibody
    • DNA helicase antibody
    • DNA helicase, RecQ like type 3 antibody
    • Exonuclease WRN antibody
    • HGNC 12791 antibody
    • OTTHUMP00000225301 antibody
    • RecQ protein-like 2 antibody
    • RecQ-like type 3 antibody
    • RecQ3 antibody
    • RECQL2 antibody
    • RECQL3 antibody
    • Werner syndrome ATP-dependent helicase antibody
    • Werner syndrome helicase antibody
    • Werner syndrome protein antibody
    • Werner syndrome, RecQ helicase like antibody
    • WRN antibody
    • WRN_HUMAN antibody
    see all

Anti-Werner's syndrome helicase WRN antibody [8H3] 图像



  • Predicted band size : 160 kDa

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: Werner's syndrome helicase WRN knockout HAP1 cell lysate (20 µg)
    Lane 3: MOLT4 cell lysate (20 µg)
    Lane 4: K562 cell lysate (20 µg)
    Lanes 1 to 4: Merged signal (red and green). Green - ab66601 observed at 170 kDa. Red - loading control, ab181602, observed at 37 kDa.
    ab66601 was shown to recognize Werner's syndrome helicase WRN when Werner's syndrome helicase WRN knockout samples were used, along with additional cross-reactive bands. Wild-type and Werner's syndrome helicase WRN knockout samples were subjected to SDS-PAGE. ab66601 and ab181602 (loading control to GAPDH) were both diluted at 1/500 and 1/10 000 and incubated overnight at 4°C. Blots were developed with goat anti-rabbit IgG (H + L) and goat anti-mouse IgG (H + L) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

  • WRN helicase is illustrated above.The helicase domain is shown in black, and the nuclear localizing signal region is hatched. Epitope sites of 4H12 and 8H3 identified by the studies with various recombinant WRN helicases (Shiratori et. al., J. Cell Biol. 144: 1-9 (1999) are also shown.WRN helicase is illustrated above.The helicase domain is shown in black, and the nuclear localizing signal region is hatched. Epitope sites of 4H12 and 8H3 identified by the studies with various recombinant WRN helicases (Shiratori et. al., J. Cell Biol. 144: 1-9 (1999) are also shown.
  • All lanes : Anti-Werner's syndrome helicase WRN antibody [8H3] (ab66601) at 1/500 dilution

    Lane 1 : Extracts from cells of normal individuals
    Lane 2 : Extracts from cells of normal individuals
    Lane 3 : Extracts from cells of normal individuals
    Lane 4 : Extracts of cells from patients with Werner syndrome
    Lane 5 : Extracts of cells from patients with Werner syndrome
    Lane 6 : Purified recombinant WRN helicase made by baculovirus technology in the insect SF9 cells. See details published in the paper by Goto et al., Human Genet 105:301-307 (1999).


    Predicted band size : 160 kDa
    Western blot analysis by ab66601 of WRN helicase protein in the EBV-transformed cells from WS (Werner syndrome) patient cells and normal individuals.
  • ICC/IF image of ab66601 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab66601, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

Anti-Werner's syndrome helicase WRN antibody [8H3] (ab66601)参考文献

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