Overview

  • Product name
    Anti-BrdU antibody [BU1/75 (ICR1)] (Alexa Fluor® 555)
    See all BrdU primary antibodies
  • Description
    Rat monoclonal [BU1/75 (ICR1)] to BrdU (Alexa Fluor® 555)
  • Host species
    Rat
  • Conjugation
    Alexa Fluor® 555. Ex: 555nm, Em: 565nm
  • Specificity
    This antibody reacts with BrdU in single stranded DNA, BrdU attached to a protein carrier or free BrdU. It detects nucleated cells in S-Phase which have had BrdU incorporated into their DNA. Also reacts with chlorodeoxyuridine but with reduced staining. The antibody does not react with thymidine. The antibody does not cross react with IdU.
  • Tested applications
    Suitable for: ICC/IFmore details
  • Species reactivity
    Reacts with: Species independent
  • Immunogen

    The details of the immunogen for this antibody are not available.

  • Positive control
    • ICC/IF: HeLa cells treated with BrdU (10uM for 24 hours)
  • General notes

    The antibody recognises single stranded DNA so the DNA needs to be unraveled first. This can be done with DNAse, although this doesn't give the best results. Depending on the assay, acid denaturation with 2M HCL or heat denaturation are the most successful. Please note this step is critical in any assay with this antibody and is the area that should be modified to optimise results. Detailed BrdU protocol is available in "Neuroscience protocols" on our "Protocol and troubleshooting tips" webpage.

    Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company. The Alexa Fluor® dye included in this product is provided under an intellectual property license from Life Technologies Corporation. As this product contains the Alexa Fluor® dye, the purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). As this product contains the Alexa Fluor® dye the sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: in manufacturing; (ii) to provide a service, information, or data in return for payment (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are sold for use in research. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, 5781 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@thermofisher.com.

Properties

Applications

Our Abpromise guarantee covers the use of ab221240 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/IF 1/100.

This product gave a positive signal in HeLa cells treated with BrdU (10uM for 24 hours) fixed with 4% formaldehyde (10 min) and 100% methanol (5 min)

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

  • ab221240 staining BrdU in HeLa cells. Untreated and BrdU treated (10uM for 24 hours) cells were fixed with 100% methanol (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 permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab221240 at 1/100 dilution (pseudocolored in green) and ab195884, Rat monoclonal to Tubulin (Alexa Fluor® 647), at 1/250 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    This product also gave a positive signal under the same testing conditions in HeLa cells fixed with 4% formaldehyde (10 min).

References

This product has been referenced in:
  • Liu L  et al. Overexpression of FoxM1 predicts poor prognosis of intrahepatic cholangiocarcinoma. Aging (Albany NY) 10:4120-4140 (2018). Read more (PubMed: 30580327) »
See 1 Publication for this product

Customer reviews and Q&As

There are currently no Customer reviews or Questions for ab221240.
Please use the links above to contact us or submit feedback about this product.

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Sign up