製品の概要

  • 製品名Anti-FEN1 antibody [4E7]
    FEN1 一次抗体 製品一覧
  • 製品の詳細
    Mouse monoclonal [4E7] to FEN1
  • アプリケーション適用あり: Flow Cyt, WB, IHC-P, ICC/IFmore details
  • 種交差性
    交差種: Human
  • 免疫原

    Recombinant human FEN-1 protein encoding amino acids 1-380 purified from baculovirus-infected Sf-9 cells.

  • ポジティブ・コントロール
    • MCF-7 cells, recombinant fusion protein.
  • 特記事項FEN-1 (Flap Endonuclease and Five prime Exonuclease) is a 42 kDa protein that functions as an endonuclease that cleaves branched DNA structures containing single-stranded 5' flaps or as an exonuclease on DNA nicks and gaps. FEN-1 endonuclease and exonuclease activities are stimulated by an association with proliferating cell nuclear antigen (PCNA) in the nucleus. FEN-1 is believed to have important roles in DNA replication and repair and is required in cell-free DNA replication systems.

製品の特性

  • 製品の状態Liquid
  • 保存方法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.
  • バッファーPreservative: None
    Constituents: PBS, pH 7.4
  • Concentration information loading...
  • 精製度Protein G purified
  • ポリ/モノモノクローナル
  • クローン名4E7
  • ミエローマNS1
  • アイソタイプIgG1
  • 軽鎖の種類kappa
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab462 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
Flow Cyt Use 0.1-1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB Use a concentration of 1 - 5 µg/ml. Predicted molecular weight: 43 kDa.
IHC-P Use a concentration of 2 µg/ml.
ICC/IF 1/500.

ターゲット情報

  • 機能Structure-specific nuclease with 5'-flap endonuclease and 5'-3' exonuclease activities involved in DNA replication and repair. During DNA replication, cleaves the 5'-overhanging flap structure that is generated by displacement synthesis when DNA polymerase encounters the 5'-end of a downstream Okazaki fragment. It enters the flap from the 5'-end and then tracks to cleave the flap base, leaving a nick for ligation. Also involved in the long patch base excision repair (LP-BER) pathway, by cleaving within the apurinic/apyrimidinic (AP) site-terminated flap. Acts as a genome stabilization factor that prevents flaps from equilibrating into structurs that lead to duplications and deletions. Also possesses 5'-3' exonuclease activity on nicked or gapped double-stranded DNA, and exhibits RNase H activity. Also involved in replication and repair of rDNA and in repairing mitochondrial DNA.
  • 配列類似性Belongs to the XPG/RAD2 endonuclease family. FEN1 subfamily.
  • 翻訳後修飾Acetylated by EP300. Acetylation inhibits both endonuclease and exonuclease activity. Acetylation also reduces DNA-binding activity but does not affect interaction with PCNA or EP300.
    Phosphorylation upon DNA damage induces relocalization to the nuclear plasma. Phosphorylation at Ser-187 by CDK2 occurs during late S-phase and results in dissociation from PCNA.
    Methylation at Arg-192 by PRMT5 impedes Ser-187 phosphorylation and increases interaction with PCNA.
  • 細胞内局在Nucleus > nucleolus. Nucleus > nucleoplasm. Mitochondrion. Resides mostly in the nucleoli and relocalizes to the nucleoplasm upon DNA damage.
  • Information by UniProt
  • 参照データベース
  • 別名
    • DNase IV antibody
    • FEN-1 antibody
    • FEN1 antibody
    • FEN1_HUMAN antibody
    • Flap endonuclease 1 antibody
    • Flap structure specific endonuclease 1 antibody
    • Flap structure-specific endonuclease 1 antibody
    • hFEN-1 antibody
    • hFEN1 antibody
    • Maturation factor 1 antibody
    • MF1 antibody
    • Rad2 antibody
    see all

Anti-FEN1 antibody [4E7] 画像

  • Overlay histogram showing HeLa cells stained with ab462 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab462, 0.1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-mouse IgG (H&L) (ab150113) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

  • ab462 detecting FEN1 (green) in Hela Cells by ICC/IF. Cells were counterstained with DAPI in order to highlight the nucleus (red). Please refer to Abreview for further experimental details.

    See Abreview

  • ab462 (2µg/ml) staining FEN1 in human duodenum sing an automated system (DAKO Autostainer Plus). Using this protocol there is strong nuclear staining .
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 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.

Anti-FEN1 antibody [4E7] (ab462) 使用論文

This product has been referenced in:
  • Qian L  et al. Human Fanconi anemia complementation group a protein stimulates the 5' flap endonuclease activity of FEN1. PLoS One 8:e82666 (2013). WB ; Human . Read more (PubMed: 24349332) »
  • Ruhanen H  et al. Involvement of DNA ligase III and ribonuclease H1 in mitochondrial DNA replication in cultured human cells. Biochim Biophys Acta 1813:2000-2007 (2011). WB ; Human . Read more (PubMed: 21878356) »

See all 7 Publications for this product

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Thank you for your enquiry and your interest in our products.

I regret to inform you that this particular batch (576831) has been sold out and we do no longer have it in our stock.

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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 Paraformaldehyde
Username

Dr. Kirk McManus

Verified customer

投稿 Oct 04 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (breast cancer cell lines)
Loading amount 3000 cells
Specification breast cancer cell lines
Blocking step Milk as blocking agent for 2 hour(s) and 30 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

投稿 Aug 16 2006

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (breast cancer cell lines)
Specification breast cancer cell lines
Username

Abcam user community

Verified customer

投稿 Aug 16 2006

The Fen-1 antibodies have not been tested in IP experiments in either mouse or human cells

You have only given the antibody's clone name. Please provide the antibody name (i.e. antigen) so I can answer with the correct information.

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