Overview

  • Product name
    Anti-BrdU antibody (HRP)
    See all BrdU primary antibodies
  • Description
    Sheep polyclonal to BrdU (HRP)
  • Host species
    Sheep
  • Conjugation
    HRP
  • Specificity
    The antibody was tested using immunoprecipitation against 5-Methyl Cytosine (5-MeC) and bromo-deoxyuridine (BrdU) or control (no antigen) and assayed by A-405 spectrophotometry. At a concentration of 25 µg/mL, this product demonstrates 8 fold higher reactivity against BrdU than 5-MeC. This product titers out at 50 ng/mL. For best results, use the product at a concentration of 25 to 100 µg/mL. Nearly complete immunoprecipitation was obtained at a concentration of 100 to 500 µg/mL. This antibody is not species specific. The nucleotide sequence is present in all species.
  • Tested applications
    Suitable for: ELISA, IHC-P, IHC-Fr, IP, WBmore details
  • Species reactivity
    Reacts with: a wide range of other species
  • Immunogen

    Bromodeoxyuridine coupled to keyhole limpet hemocyanin (KLH)

Properties

Applications

Our Abpromise guarantee covers the use of ab2285 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
ELISA Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
WB Use at an assay dependent concentration.

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

References

This product has been referenced in:
  • Sano H  et al. Identification of 5-methylcytosine in DNA fragments immobilized on nitrocellulose paper. Proc Natl Acad Sci U S A 77:3581-5 (1980). Read more (PubMed: 6251470) »
See 1 Publication for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A

Answer

Thank you for your inquiry. Yes, I can confirm that you are absolutely right, you will just need the DAB.

Indeed, HRP is the enzyme (horse radish peroxidase) which will convert the substrate (DAB) into the colored precipitation seen on the slide. The antibody ab2285 contains already HRP molecules coupled to it.

Streptavidin-HRP is need for biotinylated antibodies. Indeed, biotin and streptavidin bind very strongly together. So if an antibody is biotinylated, streptavidin coupled to HRPcan be used to bring the enzyme to the antibody/biotin.

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Question
Answer

The laboratory has used this antibody at a concentration of 10 µg/mL to obtain a good signal, comparable to other antibodies. As the antibody is at a concentration of 0.5mg/ml, this corresponds to a dilution of 1/50.

We do however recommend that the Endusers do optimise the the dilutions/concentrations, as depending on the tissues and the reagents, the optimal working dilution/concentration can vary.

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Question
Answer

The actual protocols used can be found in the following publication. Campana D, Coustan-Smith E., Janossy G., Department of Immunology, Royal Free Hospital School of Medicine, London, UK. Double and triple staining methods for studying the proliferative activity of human B and T lymphoid cells. J Immunol Methods 1071):79-88 1988 Feb 24 "ABSTRACT New methods for double and triple colour labeling using monoclonal antibodies to the proliferation-associated markers 5-methly-cytosine, BrdU and Ki67 are described. In order to make incorporated 5-methyl-cytosine or BrdU accessible to most antibodies, mild denaturation of the DNA is needed, and this is usually obtained by exposing the cells to acid or base. This procedure destroys most cellular antigen, including nuclear TdT and Ki67. In this study, we show that fixation in cold methanol instead of 70% ethanol 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. This method is optimal for detection of nuclear Ki67 and TdT. These reagents, together with antibodies to membrane assigns can be included in triple colour labeling using second layers conjugated to FITC, TRITC and colloidal gold. With these methods it is now possible to characterize the phenotype of dividing cell populations such as precursors in central lymphoid tissues and germinal centre blasts in peripheral lymphoid organs. "

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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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