Anti-Werner's syndrome helicase WRN抗体(ab200)

概述

  • 产品名称Anti-Werner's syndrome helicase WRN抗体
    参阅全部 Werner's syndrome helicase WRN 一抗
  • 描述
    兔多克隆抗体to Werner's syndrome helicase WRN
  • 特异性This antibody is specific for Werner's syndrome helicase. In Western blot the antibody detects a band at ~160 kDa representing Werner's syndrome helicase, not recognised in human fibroblast WRN knock-out cells. Works well in immunofluoresence on WRN +/+ cells and is negative on WRN -/- cells.
  • 经测试应用适用于: ELISA, ICC, ICC/IF, WBmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Synthetic peptide, corresponding to amino acids 1-510 of Human Werner's syndrome helicase WRN

  • 阳性对照
    • WRN +/+ cells

性能

应用

Our Abpromise guarantee covers the use of ab200 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
ELISA Use at an assay dependent dilution. PubMed: 17118963
ICC Use at an assay dependent dilution.
ICC/IF Use at an assay dependent concentration.
WB 1/2000. Predicted molecular weight: 162 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 图像

  • ab200 (1/1000) staining Werner's syndrome helicase WRN in HeLa cells (green). Cells were fixed in methanol, permeabilized with 0.5% Triton X100/ PBS and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

    See Abreview

  • ab200 (4µg/ml) staining Werner's syndrome helicase WRN in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is nuclear and some cytoplasmic staining.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH 6.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • All lanes : Anti-Werner's syndrome helicase WRN antibody (ab200) at 1/5000 dilution

    Lane 1 : Human primary fibroblasts whole cell lysate
    Lane 2 : Human primary fibroblasts whole cell lysate - siRNA against WRN

    Lysates/proteins at 30 µg per lane.

    Secondary
    Alexa Fluor® 680-conjugated goat anti-rabbit IgG at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 162 kDa
    Observed band size : 160 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    This image is courtesy of an anonymous Abreview

    Blocked using LI-COR® Odyssey® Blocking Buffer for 45 minutes at room temperature.

    Primary antibody dilution buffer: Blocker 1:1 in PBS + 0.1% Tween.

    See Abreview

Anti-Werner's syndrome helicase WRN antibody (ab200)参考文献

This product has been referenced in:
  • Silva BA  et al. DNA damage to a single chromosome end delays anaphase onset. J Biol Chem 289:22771-84 (2014). ICC/IF . Read more (PubMed: 24982423) »
  • Barefield C & Karlseder J The BLM helicase contributes to telomere maintenance through processing of late-replicating intermediate structures. Nucleic Acids Res 40:7358-67 (2012). WB ; Human . Read more (PubMed: 22576367) »

See all 14 Publications for this product

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (4-16% gradient Tris-Gly gel)
Sample Mouse Cell lysate - whole cell (MEFs)
Specification MEFs
Blocking step LI-COR­ Odyssey­ Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Mar 17 2015

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (4-16% gradient Tris-Gly gel)
Sample Human Cell lysate - whole cell (Human primary fibroblasts)
Specification Human primary fibroblasts
Blocking step LI-COR­ Odyssey­ Blocking Buffer as blocking agent for 45 minute(s) · Concentration: 50% · Temperature: RT°C
Username

Abcam user community

Verified customer

提交于 Mar 16 2015

Thank you for your reply.
Your credit note ID is ********.
I am sorry that these antibodies did not perform as stated on the datasheet. I have asked our accounting department to issue a credit note for you, which can be redeemed against the i...

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Thank you for your reply.
There will be a delay in you receiving these antibodies as ab2179 is currently out of stock, but it is due to come in on June 11th and we will hopefully send it to you the same day. I am sorry about the delay.
Your n...

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Thank you for calling Abcam earlier today.
I am sorry about the difficulty you have been having with a few of our antibodies and below are links to a few that would make a suitable alternative:
- http://www.abcam.com/Werner-s-syndrome-helicas...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Methanol
Permeabilization Yes - 0.5% Triton X100 in PBS
Username

Dr. Kirk McManus

Verified customer

提交于 Oct 30 2007

Thank you for your enquiry. You are correct the publication that you refer to; Agrelo R et al. Epigenetic inactivation of the premature aging Werner syndrome gene in human cancer. Proc Natl Acad Sci U S A 103:8822-7 (2006). PubMed: 16723399 ap...

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Thank you for your enquiry. The immunogen used to make this antibody was the N-terminus, amino acids 1-510, of the protein. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Thank you for your enquiry and for taking your time to fill in the on-line questionnaire. We are very sorry to her that your customer is having problem with this antibody. Would you be so kind to provide further information about: 1. How much pr...

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N-terminal of WRN. Aminoacids 1- 510

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