製品の概要

  • 製品名
  • 製品の詳細
    Rabbit polyclonal to DNA2
  • アプリケーション
    適用あり: WB, IHC-Pmore details
  • 種交差性
    交差種: Human
    交差が予測される動物種: Mouse, Rat
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 1000 to the C-terminus of Human DNA2.

    (Peptide available as ab42548.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following Human Whole Cell lysates; HEK293, HEK293 (Nuclear) and U2OS and FFPE human liver tissue sections.
  • 特記事項
    ab96488 is raised using the same immunogen as ab42439. Batches sold under ab96488 have passed QC testing in Western Blot. In our hands, this product did not work in ICC/IF. For further information please contact our Customer Services team.

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: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • 精製度
    Immunogen affinity purified
  • ポリ/モノ
    ポリクローナル
  • アイソタイプ
    IgG
  • 研究分野

アプリケーション

Our Abpromise guarantee covers the use of ab96488 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
WB Use a concentration of 1 µg/ml. Detects a band of approximately 125 kDa (predicted molecular weight: 120 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

ターゲット情報

画像

  • All lanes : Anti-DNA2 antibody (ab96488) at 1 µg/ml

    Lane 1 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
    Lane 2 : HEK293 (Human embryonic kidney cell line) Nuclear Lysate
    Lane 3 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 120 kDa
    Observed band size : 125 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 16 kDa,18 kDa,210 kDa,37 kDa,38 kDa,54 kDa,60 kDa,70 kDa,76 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 10 minutesThis antibody detects multiple bands in addition to the band of interest, reflecting low levels of expression of DNA2. siRNA knockdown experiments have demonstrated that this antibody detects a band at ~120 kDa corresponding to human DNA2 (please see image courtesy of Julien Duxin).
  • All lanes : Anti-DNA2 antibody (ab96488)

    Lane 1 : U2OS cells plus control shRNA
    Lane 2 : U2OS cells plus shDNA2-1 shRNA
    Lane 3 : U2OS cells plus shDNA2-2 shRNA

    Secondary
    Anti-rabbit at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 120 kDa
    Observed band size : 120 kDa

    Image courtesy of Julien Duxin and Sheila Stewart

    Western blot showing detection of DNA2 by ab42439. A 120 kDa band corresponding to DNA2 was detected in lysates of U2OS cells treated with control shRNA (lane 1) but not in those from knockdown cells treated with two different shRNAs for DNA2 (lanes 2 and 3). Following pre-blocking for 30 mins in 5% milk, PBS-Tween, the membrane was incubated overnight at 4°C with ab42439 at a dilution of 1/1000 in 5% milk, PBS-Tween. The membrane was then incubated with the secondary antibody at 1/3000 dilution in 5% milk, PBS-Tween for 2 hours at RT, and washed 3 x 10 mins with PBS-Tween prior to ECL detection.
  • IHC image of ab96488 staining in human liver carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab96488, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

参考文献

This product has been referenced in:
  • Gong Y  et al. PHF11 promotes DSB resection, ATR signaling, and HR. Genes Dev 31:46-58 (2017). WB ; Mouse . Read more (PubMed: 28115467) »
  • Biehs R  et al. DNA Double-Strand Break Resection Occurs during Non-homologous End Joining in G1 but Is Distinct from Resection during Homologous Recombination. Mol Cell 65:671-684.e5 (2017). Read more (PubMed: 28132842) »

See all 12 Publications for this product

レビューと Q&A

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (U2OS cells)
Total protein in input
500 µg
Immuno-precipitation step
Protein A/G
Specification
U2OS cells
Username

Abcam user community

Verified customer

投稿 Jul 18 2017

Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
50 µg
Specification
U2OS cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Oct 26 2016

Thank you for contacting us. Unfortunately, this antibody does not appear to be appropriate for ICC/IF. The Abreview showed poor results for this application and the in house testing has confirmed that the antibody should not be recommended for...

Read More
Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Specification
HeLa
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.5% Triton X100
Username

Dr. Kirk Mcmanus

Verified customer

投稿 Dec 09 2010

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

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