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  1. Link

    brdu-cell-proliferation-elisa-kit-colorimetric-ab126556.pdf

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BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

  • Datasheet
  • SDS
  • Protocol Booklet
Reviews (2)Q&A (5)References (114)

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Indirect ELISA - ab126556 BrdU Cell Proliferation ELISA Kit (colorimetric)
  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

Key features and details

  • Sensitivity: 40 cells/well
  • Sample type: Adherent cells, Suspension cells
  • Detection method: Colorimetric
  • Assay type: Sandwich (qualitative)

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Overview

  • Product name

    BrdU Cell Proliferation ELISA Kit (colorimetric)
    See all BrdU kits
  • Detection method

    Colorimetric
  • 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

    BrDU Cell proliferation ELISA kit (ab126556) is an indirect ELISA kit for the detection of BrdU incorporation into newly synthesized DNA of actively proliferating cells. It 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.


    Store kit at +4°C immediately upon receipt except for the Prediluted Anti-BrdU Detector Antibody and Peroxidase Goat anti-mouse IgG (2000X) which must be stored at -20°C.

  • Notes

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • Platform

    Microplate

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 200 tests
    500X BrdU Reagent 1 x 15µl
    Conjugate Diluent 1 x 25ml
    Fixing Solution 2 x 20ml
    Peroxidase Goat anti-mouse IgG (2000X) 1 x 15µl
    Plate wash concentrate (50X) 1 x 90ml
    Prediluted anti-BrdU detecting antibody 1 x 20ml
    Stop Solution 1 x 25ml
    TMB Substrate 1 x 25ml
  • Research areas

    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Cell cycle proteins ELISA kits
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Fusion/marker protein ELISA Kits

Associated products

  • Related Products

    • Anti-BrdU antibody [BU1/75 (ICR1)] - Proliferation Marker (ab6326)

Images

  • Indirect ELISA - ab126556 BrdU Cell Proliferation ELISA Kit (colorimetric)
    Indirect ELISA - ab126556 BrdU Cell Proliferation ELISA Kit (colorimetric)

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

  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
    Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

    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.

  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
    Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

    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.

  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
    Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

    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.

  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
    Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

    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.

  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
    Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

    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.

  • Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)
    Indirect ELISA - BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

    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.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (114)

Publishing research using ab126556? Please let us know so that we can cite the reference in this datasheet.

ab126556 has been referenced in 114 publications.

  • Bhansali S  et al. Hypoxia-induced mitochondrial reactive oxygen species (mtROS) differentially regulates smooth muscle cell (SMC) proliferation of pulmonary and systemic vasculature. Mitochondrion 57:97-107 (2021). PubMed: 33253916
  • Shou J  et al. LncRNA HOXA-AS2 promotes glioblastoma carcinogenesis by targeting miR-885-5p/RBBP4 axis. Cancer Cell Int 21:39 (2021). PubMed: 33430870
  • Küper A  et al. Overcoming hypoxia-induced resistance of pancreatic and lung tumor cells by disrupting the PERK-NRF2-HIF-axis. Cell Death Dis 12:82 (2021). PubMed: 33441543
  • Chen H  et al. miR-27a-3p regulates the inhibitory influence of endothelin 3 on the tumorigenesis of papillary thyroid cancer cells. Mol Med Rep 23:N/A (2021). PubMed: 33537832
  • Luo C  et al. MicroRNA-640 promotes cell proliferation and adhesion in glioblastoma by targeting Slit guidance ligand 1. Oncol Lett 21:161 (2021). PubMed: 33552279
View all Publications for this product

Customer reviews and Q&As

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1-7 of 7 Abreviews or Q&A

Cell Proliferation assay in MCF 7 cells

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Followed the same protocol as per booklet..
Worked well
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Mar 30 2018

Fast protocol, easy to use, though certain limitations

Good Average 3/5 (Ease of Use)
Abreviews
Abreviews
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.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jul 10 2017

Question


Inquiry: I am going to perform the BrdU assay on cells which are adhesive but are also quite fragile. With other similar assays I have been having trouble with variability between replicates because cells get easily detached through the process. I was wondering if I could use DAPI as a means to normalise the BrdU signal to the number of cells still present in each well (whether these are dead or alive)? Do you have a suggestions of how this would be acheived?

Read More

Abcam community

Verified customer

Asked on Apr 11 2016

Answer

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

Abcam Scientific Support

Answered on Apr 11 2016

Question

I have just bought the following item from your company : BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556).

I would like to normalize each well containing BrdU to the number of total live cells. What method would you advise me to use? I was thinking to normalize to the total number of protein or of DNA content but I can't see how these methods can be combined with the BrdU since the cells are immunostained and fixed.















Read More

Abcam community

Verified customer

Asked on Aug 13 2014

Answer

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.

Read More

Abcam Scientific Support

Answered on Aug 13 2014

Question

I have some questions regarding the protocol for ab126556:
1. Is 20ul Brdu, as described in the protocol, added to each well regardless of whether the well contains 100 or 200ul?
2. How are the results to be calculated?
3. Please explain the data showed on the example data included in the protocol.
4. Why is it suggested to either measure OD450-OD550, OD450-OD595 or OD450 alone when measuring the absorbance in each well?

Read More

Abcam community

Verified customer

Asked on Jun 05 2014

Answer



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.

Read More

Jeremy Kasanov

Abcam Scientific Support

Answered on Jun 05 2014

Question

Kindly confirm whether below kit can be used for Rat cells (Reactivity in Rat):

Read More

Abcam community

Verified customer

Asked on Jan 15 2014

Answer

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

Read More

Abcam Scientific Support

Answered on Jan 15 2014

Question

Can this be used on Pig cells?

Read More

Abcam community

Verified customer

Asked on Oct 10 2013

Answer



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

Read More

Jeremy Kasanov

Abcam Scientific Support

Answered on Oct 10 2013

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