製品の概要

  • 製品名
    Anti-Werner's syndrome helicase WRN antibody
    Werner's syndrome helicase WRN 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Werner's syndrome helicase WRN
  • 由来種
    Rabbit
  • アプリケーション
    適用あり: ELISA, ICC, IHC-P, ICC/IF, WBmore details
  • 種交差性
    交差種: Human
  • 免疫原

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

  • ポジティブ・コントロール
    • WRN +/+ cells

製品の特性

アプリケーション

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.

アプリケーション Abreviews 特記事項
ELISA Use at an assay dependent concentration. PubMed: 17118963
ICC Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB 1/2000. Predicted molecular weight: 162 kDa.

ターゲット情報

  • 機能
    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.
  • 関連疾患
    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].
  • 配列類似性
    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.
  • 翻訳後修飾
    Phosphorylated by PRKDC. Phosphorylated upon DNA damage, probably by ATM or ATR.
  • 細胞内局在
    Nucleus > nucleolus. Nucleus.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

画像

  • 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
    All lanes : 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


    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

  • 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.

参考文献

This product has been referenced in:
See all 20 Publications for this product

レビューと Q&A

1-10 of 15 Abreviews or Q&A

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HeLa)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
HeLa
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

投稿 Apr 13 2017

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

Abcam user community

Verified customer

投稿 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

Abcam user community

Verified customer

投稿 Mar 16 2015

Answer

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 invoice of a future order by passing it on to your purchasing department. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used. If you have questions on how to use the credit note, please contact our accounting department.
Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.
The credit note ID is for your reference only and does not automatically guarantee the credit.
I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

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Question
Answer

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 new order number for tracking purposes is Order ************.
If there is anything else I can help you with, please let me know.

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Question
Answer

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:
- https://www.abcam.com/Werner-s-syndrome-helicase-WRN-antibody-ab17987.html
- https://www.abcam.com/Werner-s-syndrome-helicase-WRN-antibody-EPR6392-ab124673.html
- https://www.abcam.com/Blooms-Syndrome-Protein-Blm-antibody-ab2179.html
- https://www.abcam.com/Blooms-Syndrome-Protein-Blm-antibody-ab86664.html
Please let me know which antibodies you would like as replacements and I will have them sent to you as soon as possible.

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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

Dr. Kirk Mcmanus

Verified customer

投稿 Oct 30 2007

Answer

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 applied this antibody in cultured cell lines using immunofluorescence in addition to immunohistochemistry on Werner's syndrome mice. In the publication within the methods section they refer to references 22 and 30 for their methodology in these two applications; Ropero, S., Setien, F., Espada, J., Fraga, M. F., Herranz, M., Asp, J., Benassi, M. S., Franchi, A., Patino, A., Ward, L. S., et al. (2004) Hum. Mol. Genet. 13, 2753–2765. Chen, L., Lee, L., Kudlow, B. A., Dos Santos, H. G., Sletvold, O., Shafeghati, Y., Botha, E. G., Garg, A., Hanson, N. B., Martin, G. M., et al. (2003) Lancet 362, 440–445. Unfortunately these two pubclications do in fact refer to; BK Kennedy, DA Barbie, M Classon, N Dyson and E Harlow, Nuclear organization of DNA replication in primary mammalian cells, Genes Dev 14 (2000), pp. 2855–2868 in their methods section. However, I have examined this publication and this does not detail the approach used; whether frozen sections or paraffin embedded sections. Therefore unfortunately I cannot update this antibody datasheet to reflect the antibody's application in this publication. For details of the methods employed I would encourage you to contact them directly using the email address detailed on the publications. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

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Answer

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.

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Answer

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 protein was loaded onto the gel? 2. What is the secondary antibody? In what species was it raised? We certainly can offer you a new vial free of charge. Please do let us know if your customer would like to test it. We look forward to hearing from you soon.

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1-10 of 15 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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