Anti-BrdU 抗体 [MoBu-1] (ab8039)

Submit a review Q&A (5)References (17)

Key features and details

  • Mouse monoclonal [MoBu-1] to BrdU
  • Suitable for: ICC, IHC-P, ICC/IF, Flow Cyt
  • Reacts with: Species independent
  • Isotype: IgG1

製品の概要

  • 製品名

    Anti-BrdU antibody [MoBu-1]
    BrdU 一次抗体 製品一覧

  • 製品の詳細

    Mouse monoclonal [MoBu-1] to BrdU

  • 由来種

    Mouse

  • 特異性

    This antibody reacts specifically with BrdU incorporated into DNA during S-phase of a cell cycle. It is useful for detecting proliferating cells by flow cytometry or immunofluorescence staining. The reaction shows a clear, nuclear confined speckled pattern. It reacts also specifically with 5-bromouridine (BrU).

  • アプリケーション

    適用あり: ICC, IHC-P, ICC/IF, Flow Cytmore details

  • 種交差性

    交差種: Species independent

  • 免疫原

    Chemical/ Small Molecule corresponding to BrdU.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab8039 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
ICC Use a concentration of 2 µg/ml.
IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 2 µg/ml.
Flow Cyt Use a concentration of 1 - 2 µg/ml.

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

ターゲット情報

  • 関連性

    The immunocytochemical detection of bromodeoxyuridine (BrdU) incorporated into DNA is a powerful tool to study the cytokinetics of normal and neoplastic cells. In vitro or in vivo labeling of tumor cells with the thymidine analogue BrdU and the subsequent detection of incorporated BrdU with specific anti-BrdU monoclonal antibodies is an accurate and comprehensive method to quantitate the degree of DNA-synthesis. BrdU is incorporated into the newly synthezised DNA of S-phase cells may provide an estimate for the fraction of cells in S-phase. Also dynamic proliferative information such as the S-phase transit rate and the potential doubling time can be obtained, by means of bivariate BrdU/DNA flow cytometric analysis.

  • 細胞内局在

    Nuclear

  • 別名

    • Bromodeoxyuridine antibody
    • BUdr antibody

画像

  • Immunocytochemistry/ Immunofluorescence - Anti-BrdU antibody [MoBu-1] (ab8039)
    Immunocytochemistry/ Immunofluorescence - Anti-BrdU antibody [MoBu-1] (ab8039)

    ICC/IF image of ab8039 stained HeLa cells, both BrdU treated (left image) and normal cells (right image). The cells were 100% methanol fixed (5 min) and then subjected to acid hydrolysis using 2M HCL in 0.1% PBS-Tween for 30 minutes at room temperature to denature the DNA. They were  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 (ab8039, 10µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) used at a 1/250 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. Positive staining can be seen in the BrdU treated cells, but not in the normal cells, demonstrating specificity for BrdU.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BrdU antibody [MoBu-1] (ab8039)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BrdU antibody [MoBu-1] (ab8039)

    IHC image of ab8039 staining, both in normal and BrdU treated rat liver formalin fixed paraffin embedded tissue sections, performed on a Leica Bond

    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.

  • Flow Cytometry - Anti-BrdU antibody [MoBu-1] (ab8039)
    Flow Cytometry - Anti-BrdU antibody [MoBu-1] (ab8039)

    Flow cytometry analysis of CEM (human acute lymphoblastic leukemia) cells labelling BrdU with ab8039 at 1 µg/mL. Goat anti-mouse IgG was used as the secondary antibody. The individual cell cycle phases (S, G1, G2/M-phase) are indicated on the figure. 

参考文献 (17)

ab8039 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab8039 は 17 報の論文で使用されています。

  • Xie X  et al. Optimization of In Vitro Culture Conditions of Sturgeon Germ Cells for Purpose of Surrogate Production. Animals (Basel) 9:N/A (2019). PubMed: 30901855
  • Niknam P  et al. Modulating proteoglycan receptor PTPs using intracellular sigma peptide improves remyelination and functional recovery in mice with demyelinated optic chiasm. Mol Cell Neurosci 99:103391 (2019). PubMed: 31276750
  • Mehta K & Laimins L Human Papillomaviruses Preferentially Recruit DNA Repair Factors to Viral Genomes for Rapid Repair and Amplification. MBio 9:N/A (2018). PubMed: 29440569
  • Zhang J  et al. Pericardial application as a new route for implanting stem-cell cardiospheres to treat myocardial infarction. J Physiol 596:2037-2054 (2018). PubMed: 29736937
  • Juhász S  et al. ATRX Promotes DNA Repair Synthesis and Sister Chromatid Exchange during Homologous Recombination. Mol Cell 71:11-24.e7 (2018). PubMed: 29937341
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1-5 of 5 Abreviews or Q&A

Question

Hello, I need to work with IdU and CldU for double staining. I found out that I can use for cldU the ab6326 that doen't cross react with IdU. Can I use ab8039 or ab8152 for IdU? I need an antibody that doesn't cross react with CldU.

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Answer

Thank you for your inquiry and for rating your experience with us.

I just wanted to follow up on your query with some additional information.

For ab8039, this antibody can be used for detection of BrdU and BrU. We have not tested it regarding other chemicals of this family.

For ab8152, this antibody is specific for DNA in which BrdU has been incorporated. In immunoassays this antibody reacts strongly with free or carrier-protein coupled BrdU but not with other nucleosides. In immuncytochemistry the antibody only recognizes BrdU in denaturated (single stranded) DNA. The BrdU antibody is 100% crossreactive with Iodo-Deoxy-Uridine (IrdU). Therfore, IdU instead of BrdU can be used in studies.

I hope this information helps. Please contact us with any other questions.

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Question

Hello, I need to work with IdU and CldU for double staining. I found out that I can use for cldU the ab6326 that doen't cross react with IdU. Can I use ab8039 or ab8152 for IdU? I need an antibody that doesn't cross react with CldU.

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Answer

We have not tested these antibodies for any cross-reactivity with IdU.

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Question

thank you very much for your email. Could you provide me with a reference for where this antibody has been used for flow cytometry?

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Answer

We know of one publication at this moment in time where this antibody was in flow cytometry (also stated in the reference tab of the datasheet):

http://www.ncbi.nlm.nih.gov/pubmed/18625847?dopt=Abstract

Please see the supplemental images (http://www.jcb.org/cgi/content/full/jcb.200712014/DC1)in the publicationfor more information.

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Question

Product code: 8039

Inquiry: Hi, I'm looking for a mouse monoclonal Brdu antibody that cross reacts with human cells (and ideally ovine too). It must be suitable for flow cytometry. Is this the one you would recomend? Thanks,

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Answer

Thank you for your inquiry.

I can confirm that ab8039 reacts specifically with BrdU incorporated into DNA during S-phase of a cell cycle.

It does not matter what species the DNA is from. If BrdU is incorporated into human or ovine DNA (or DNA from any other species), this antibody is suitable for you experiments in flow cytometry.

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Question

Do all antibodies to BrdU demand denaturation of DNA i.e. antibodies only attach into single-stranded DNA? Thanks! Anna Rissanen

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Answer

Yes. Fixation in cold methanol for 30 minutes followed by immersion in 7 x 10-3 N NaOH for 10-15 seconds allows BrdU staining with the simultaneous detection of nuclear cytoplasmic and membrane assigns as well as preservation of morphological detail.

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