BrdU Cell Proliferation ELISA Kit (colorimetric) (ab126556)

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

    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
    Microplate

Properties

Associated products

Applications

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.

Images

  • 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.

Protocols

References

This product has been referenced in:
  • Matta A  et al. Molecular Therapy for Degenerative Disc Disease: Clues from Secretome Analysis of the Notochordal Cell-Rich Nucleus Pulposus. Sci Rep 7:45623 (2017). Read more (PubMed: 28358123) »
  • Yi J  et al. Overexpression of NSUN2 by DNA hypomethylation is associated with metastatic progression in human breast cancer. Oncotarget 8:20751-20765 (2017). Human . Read more (PubMed: 27447970) »

See all 20 Publications for this product

Customer reviews and Q&As

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.
Username

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.  

T...

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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...

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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 ...

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



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

Thank you for the additional information. I have arranged for this replacement vial of Peroxidase Goat anti-mouse IgG (2000X) to be shipped. You should receive it by the end of next week. For your records, the order reference number associated with thi...

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Thank you for your email and I apologize for the delay in my reply.

I have aquired a new vial of Peroxidase Goat anti-mouse IgG (2000X) that I'd like to send. So I can process this shipment, please provide the PO number or Abcam order referen...

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Thank you for your enquiry.

I just want to confirm that you have centrifuged the vial of Peroxidase Goat anti-mouse IgG (2000X) to ensure any droplets potentially on the sides and in the lid have been recovered.

Thanks in advance fo...

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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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