We enable nucleic acid labeling bioconjugation
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EdU-Click 555

EdU Cell Proliferation Kit for Imaging

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  • Size
  • Catalog No.
  • Price
  • 1 Kit
  • BCK-EdU555

  • 385 €
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  • All EdU based proliferation kits on the market are patented by our company since 2007. This click assay shows several advantages over other cell viability assays (BrdU or 3H-thymidine) making it a superior evaluation tool for monitoring genotoxicity, evaluation of anticancer drugs or cell cycle analysis and for assessing cell vitality. Due to direct measurement of cell proliferation our assay is thus highly reliable and showing no toxic effects in animals. A differentiation between cells that are actively dividing and those that are quiescent is herewith possible. Measured are only the cells in vitro or in vivo that are actively dividing. Healthy cells that are not dividing are not detected and can thus give no false positives. The patented technique is based on the incorporation of the thymidine analogue EdU, an alkyne modified nucleoside, in DNA during active cell synthesis in whole animals. After incorporation, EdU is detected via click chemistry in different organs. In healthy mice, cell proliferation is nicely detected in lymph nodes, but not in other organs showing the reliability of this baseclick method.

    LITERATURE

    Salic et al., Biotechniques 2008, 44, 929-929

    Ayaydin et al., Plant Methods 2010, 6, 5

    Chapman et al., Dev. Dyn. 2009, 238, 944-949

    FAQ

    • 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).

    • Shelf Life

      12 months unopened after receipt

    • Storage Conditions

      2-8°C

    • Physical State

      kit system made of different components

    • CAS Number

      n.a.

    • Absorption (max)

      546 nm

    • Emission (max)

      579 nm

    • Ɛ (max)

      91.000 cm-1M-1

    • Preparation/Handling

      please see user manual of the kit