Overview

  • Product name
    Anti-BrdU antibody [MoBu-1]
    See all BrdU primary antibodies
  • Description
    Mouse monoclonal [MoBu-1] to BrdU
  • Host species
    Mouse
  • Specificity
    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).
  • Tested applications
    Suitable for: ICC, IHC-P, ICC/IF, Flow Cytmore details
  • Immunogen

    Chemical/ Small Molecule corresponding to BrdU.

Properties

Applications

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.

Application Abreviews Notes
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.

Target

  • Relevance
    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.
  • Cellular localization
    Nuclear
  • Alternative names
    • Bromodeoxyuridine antibody
    • BUdr antibody

Images

  • 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.

  • 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 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. 

References

This product has been referenced in:
  • Mehta K & Laimins L Human Papillomaviruses Preferentially Recruit DNA Repair Factors to Viral Genomes for Rapid Repair and Amplification. MBio 9:N/A (2018). Read more (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). Read more (PubMed: 29736937) »
See all 14 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

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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Answer

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

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

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