• Product name
    BrdU Cell Proliferation ELISA Kit (colorimetric)
    See all BrdU kits
  • Detection method
  • Sample type
    Adherent cells, Suspension cells
  • Assay type
    Sandwich (qualitative)
  • Sensitivity
    < 40 cells/well
  • Assay time
    2h 30m
  • Assay duration
    Multiple steps standard assay
  • Product overview

    ab126556 involves incorporation of BrdU into cells cultured in microtiter plates using the cell layer as the solid phase. During the final 2 to 24 hours of culture BrdU is added to wells of the microtiter plate. BrdU will be incorporated into the DNA of dividing cells. To enable antibody binding to the incorporated BrdU cells must be fixed, permeabilized and the DNA denatured. This is all done in one step by treatment with Fixing Solution. Detector anti-BrdU monoclonal antibody is pipetted into the wells and allowed to incubate for one hour, during which time it binds to any incorporated BrdU. Unbound antibody is washed away and horseradish peroxidase-conjugated goat anti-mouse antibody is added, which binds to the Detector Antibody.

    The horseradish peroxidase catalyzes the conversion of the chromogenic substrate tetra-methylbenzidine (TMB) from a colorless solution to a blue solution (or yellow after the addition of stopping reagent), the intensity of which is proportional to the amount of incorporated BrdU in the cells. The colored reaction product is quantified using a spectrophotometer.

    The resultant assay is sensitive, rapid, easy to perform and applicable to high sample throughput. In addition to evaluation of cell proliferation, information such as cell number, morphology and analysis of cellular antigens can be obtained from a single culture.

  • Tested applications
    Suitable for: Indirect ELISAmore details
  • Platform


Associated products


Our Abpromise guarantee covers the use of ab126556 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
Indirect ELISA Use at an assay dependent concentration.


  • Cell proliferation measured in fibroblasts showing cell amounts vs. optical densities

  • Detection of RH7777 (adherent) cells per well with 2 hour pulse with BrdU. Y axis - left, OD 450-550 nm. Y axis-right, signal -to-noise ratio.
  • Detection of RH7777 (adherent) cells per well with 24 hour pulse with BrdU. Y axis - left, OD 450-550 nm. Y axis-right, signal-to-noise ratio.
  • Detection of Jurkat cells (non-adherent) per well with 2 hour pulse with BrdU. Y axis - left, OD 450-550 nm. Y axis-right, signal-to-noise ratio.
  • Detection of Jurkat cells (non-adherent) per well with 24 hour pulse with BrdU. Y axis - left, OD 450-550 nm. Y axis-right, signal-to-noise ratio.

  • Detection of MCF7 cells per well with 2 hour pulse with BrdU. Y axis - left, OD 450-550 nm. Y axis-right, signal-to-noise ratio.
  • Detection of MCF7 cells per well with 24 hour pulse with BrdU. Y axis - left, OD 450-550 nm. Y axis-right, signal-to-noise ratio.



This product has been referenced in:
  • Hoque SAM  et al. Mitochondrial Protein Turnover Is Critical for Granulosa Cell Proliferation and Differentiation in Antral Follicles. J Endocr Soc 3:324-339 (2019). Read more (PubMed: 30652133) »
  • Di Dalmazi G  et al. MYMD-1, a Novel Immunometabolic Regulator, Ameliorates Autoimmune Thyroiditis via Suppression of Th1 Responses and TNF-a Release. J Immunol 202:1350-1362 (2019). Read more (PubMed: 30674573) »
See all 60 Publications for this product

Customer reviews and Q&As

1-7 of 7 Abreviews or Q&A

Cell Proliferation assay in MCF 7 cells

Excellent Excellent 5/5 (Ease of Use)
Followed the same protocol as per booklet..
Worked well

Abcam user community

Verified customer

Submitted Mar 30 2018

This BrdU assay offers a quick and reliable approach to the assessment of cellular proliferation. However, the prediluted primary antibody (20 ml) will not suffice for 200 tests if you use a multichannel pipette due to pipetting errors.

Abcam user community

Verified customer

Submitted Jul 10 2017


The fixative solution provided with BrdU Cell Proliferation ELISA Kit (ab126556) does an excellent job of keeping the cells in the wells.  So it should not be necessary to do anything further to normalize the signal to the number of cells.  

That being said, it is an interesting idea to do it.  If the reader can detect DAPI, it might be possible to test the DAPI with the fixing solution and substrate/stop solutions provided with the kit.  The largest unknown would be how the DAPI will react with the addition of the substrate and stop solution.  The substrate can be read without the stop solution, if necessary, (at 650 nm) but it will lose sensitivity.  

So if you have the ability to read the DAPI and the TMB signals, you can can try it.  However, this may not work and we cannot guarantee it. Therefore, if you want to try, we would recommend to build controls into the assay design and also know the number of cells that have been plated.  Bad CVs can be dealt with using an increased number of replicates.  

Read More


If we understand your request correctly, you are trying to control to assure an equal number of viable cells is present in each assay well plus and minus BrdU. Our only suggestion would be for you to perform the assay in test tubes or deep well 96-well culture plates and count the viable cells prior to a short (˜2 hour) BrdU incubation. Alternatively, duplicate plates can be set up and one set can be assessed for cell number and one used for BrdU. Checking for protein or DNA content will not give an accurate cell viability count. Dead cells in the wells will have just as much protein and DNA as viable cells.

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Q1: Is 20ul Brdu, as described in the protocol, added to each well regardless of whether the well contains 100 or 200ul?
A1: Yes. The amount of Brdu is in excess, so it is fine to add the same 20ul of Brdu whether or not you have added a test reagent. However if you do add a test reagent please ensure that every well has 200ul medium as a final volume, even if not every well contains a test reagent. This is becuase more medium in a well will provide more nutrients for the cells, causing them to perhaps proliferate faster or to a greater degree, throwing off your data.

Q2: How are the results to be calculated?
A2: For each experiment, you will have a well treated with Brdu or not treated with Brdu. Just subtract the wells without Brdu from those which were treated with Brdu. -Brdu wells serve as internal control for background. You can run a blank control as mentioned in the protocol as well. However this is just to ensure that there is nothing inherently wrong with the assay. This blank should always have a very low OD such that subtracting it from all wells will have no effect on the data. However if it is a high value then it is a good indicator there is a problem with the assay.

Q3: Please explain the data showed on the example data included in the protocol.
A3: The green bars should be measured with the Y axis on the right. It measures the ratio of wells treated with and without Brdu (+Brdu/-Brdu). The curves with black and white circles should be qantified using the Y axis on the left. The black circles are OD values from wells treated with Brdu and white circles are from corresponding control wells not treated with Brdu.

Q4: Why is it suggested to either measure OD450-OD550, OD450-OD595 or OD450 alone when measuring the absorbance in each well?
A4: OD550 or OD595 readings act as the reference wavelength. A reference wavelength is used to take into account any variations in the data due to the spec itself. However this reference wavelength value is typically very small so not always used.

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Your customer can use this product on rat cells, as it has no species restriction

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Yes, ab126556 can be used with pig cells. This kit detects incorporation of BrdU independent of the species of the cell line being tested.

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