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Cell Proliferation

EdU Cell Proliferation Assay Kits

baseclick GmbH

A superior cell proliferation assay

Do you need to stop working with 3H-thymidine for cell proliferation analysis? Are you looking for a sensitive and reliable cell proliferation assay? We have developed a cell proliferation assay with a simple workflow for detection by fluorescence microscopy, flow cytometry and for high-throughput screening in fluorescence plate readers. Moreover, baseclick´s EdU cell proliferation kits are available for cell proliferation assessment in vivo.

Most recently, we have developed EdU DetectPro Kits, which possess an even improved sensitivity. Due to our patented technology we provide superior cell proliferation kits at a reasonable price.

The key attributes are:

  • A fast detection procedure (30 min) to save your time
  • Cell proliferation detection at affordable cost to save your money
  • No radioactive compounds are needed to protect your health and the environment
  • Highly reliable and efficient method without false positives
  • Mild protocol conditions without DNA denaturation to preserve your samples and allow for multiplexing
  • More sensitive cell proliferation detection compared to BrdU kits for deeper insights

How do the kits work?

Baseclick EdU cell proliferation kits are based on the incorporation of 5-ethynyl deoxyuridine, abbreviated as EdU, which is an alkyne-modified thymidine analogue. EdU is taken up by the host cell, gets phosphorylated and is incorporated into DNA during the S-phase of the cell cycle. After EdU incorporation, fixation and permeabilization of the sample, de novo synthesized DNA is detected via click chemistry with fluorescent dyes.

Applications

Baseclick´s cell proliferation kits can be used to determine the influence of (small) molecules, chemicals and drugs on DNA synthesis to evaluate e.g. genotoxicity in oncology, REACH testing and for drug substance screening.

Which kit is the right for me?

Baseclick EdU cell proliferation kits for imaging are designed for fluorescence microscopy detection and are offered for different readout filter settings. This allows adaption to your detection setup and enables simultaneous detection of additional parameter(s) for multiplexing.

EdU cell proliferation kits for flow cytometry are most likely the best way to quantify cell proliferation by measuring new DNA synthesis and provide options for automated data acquisition.
EdU cell proliferation kits for high-throughput screening (HTS) provide reagents for multiple microtiter plates and can be adapted for automation. Please note that only a limited amount of fluorescent dyes is available for the HTS format.
Baseclick EdU in vivo kits contain an increased amount of EdU for feeding in an animal or plant model organism and are available for imaging, flow cytometry and HTS.

Do you need more information on EdU cell proliferation kits? See our FAQs.
Are you interested to purchase a cell proliferation kit? Request a quote!

Literature

Cell Proliferation

  • EdU for cell proliferation, a more efficient alternative to BrdU and H3dU[1]
  • Flow Cytometry analysis of cell proliferation based on EdU[2][3]
  • EdU cell proliferation in nervous system[4]
  • Immunoproliferative response analysis using EdU[5]

Selected references

[1] P. Cappella, F. Gasparri, M. Pulici, J. Moll, Cytom. Part A 2008, 73, 626–636.
[2] Y. Sun, Y. Sun, G. Lin, R. Zhang, K. Zhang, J. Xie, L. Wang, J. Li, Cytom. Part A 2012, 81 A, 901–909.
[3] S. Diermeier-Daucher, S. T. Clarke, D. Hill, A. Vollmann-Zwerenz, J. A. Bradford, G. Brockhoff, Cytom. Part A 2009, 75, 535–546.
[4] C. Zeng, F. Pan, L. a Jones, M. M. Lim, E. a Griffin, Y. I. Sheline, M. a Mintun, D. M. Holtzman, R. H. Mach, 2011, 21–32.
[5] Y. Kitazawa, H. Ueta, T. Hünig, Y. Sawanobori, K. Matsuno, Histochem. Cell Biol. 2015, 144, 195–208.
Jungmann, Nat. Methods 2018, 15, DOI 10.1038/s41592-018-0105-0.

FAQs

  • What type of cells can incorporate EdU?

    The EdU cell proliferation assay has been applied to many different cell types and organism. Cancer cell lines like HeLa, HEK, MOLM are arguably among the most routine applications, but also animals, like mouse, rat, the nematode C. elegans, crickets (Gryllus bimaculatus), chicken (Gallus domesticus) and zebra fish (Danio rerio) or even plants (e.g. Arabidopsis thaliana) can be applied.

    Cells that possess a pyrimidine salvage pathway can phosphorylate EdU to the corresponding triphosphate, which is then accepted by the host DNA polymerase for incorporation into DNA during replication and therefore they are compatible with the EdU assay.

  • Can I perform EdU cell proliferation detection on live cells?

    No, at the moment the kit needs live cells to incorporate the EdU, but the detection reaction must be performed on fixed and permeabilized samples. The click cocktail is cell impermeant.

  • How does EdU labeling compare to BrdU or the 3H-thymidine incorporation assay?

    All three methods enable to determine cell proliferation directly, by incorporation of a metabolite analogue and subsequent detection. The 3H-thymidine incorporation assay is very sensitive, but the radioactive compound requires specialized equipment and dedicated lab space for handling.

    EdU and BrdU assays are non-radioactive alternatives with decreased risk for health and environment. Compared to the BrdU incorporation assay the EdU assay is more sensitive, requires less handling time and needs no harsh DNA denaturing conditions for detection. Therefore, the EdU cell proliferation is also compatible with multiplexing.

  • Can I combine DAPI staining and EdU detection?

    Yes, this is feasible. Please note that DAPI staining should be done after the click detection step. Alternatively, SYBR Green DNA staining can be used. But, please note that SYBR green is not compatible with the 488 EdU kits.

  • When can I stop safely the protocol?

    It is possible to safely stop the protocol after the fixation step, remove the fixation solution and wash as suggested by the user manual, then the cells can be stored in buffer at 4° C, the permeabilization step is not required immediately. The protocol can be safely stopped even after the permeabilization, following the same recommendation as before.

    It is important to not stop the protocol if the click cocktail for the detection of the EdU has been prepared, since the cocktail reacts optimally within 15 minutes.

  • Is antibody staining compatible with the EdU?

    Antibody staining is compatible with EdU cell proliferation detection when antibody detection is done after the click detection step. Please make sure that the detection signals of both detection methods do not overlap due to identical or similar spectral properties of the dyes.

    Check also the user manual for more information.

  • What is the right duration of EdU incubation?

    The duration of the EdU incubation step depends on your cell type or organism, the applied EdU concentration and even the experimental design. For a start it is advisable to refer to a published protocol (which is close to your experimental setup) and to test the conditions with a low number of samples.

    As a general guideline we recommend to use a maximum of 10 µM final EdU in the cell culture medium for incubations of a few hours to maximum one day. For longer incubation durations the concentration should be decreased to 1-5 µM. Cells that divide rapidly (once a day) generally require shorter incubation compared to cells with slow division (once a week).

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