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ClickTech EdU Cell Proliferation Kit 555 (50 Assays) for FC IV

In vivo EdU Cell Proliferation Kit for Flow Cytometry

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  • Size
  • Catalog No.
  • Price
  • 1 Kit size S (+50 mg EdU)
  • BCK-EdU555FC50+IV-S

  • € 525,00
  • 1 Kit size M (+500 mg EdU)
  • BCK-EdU555FC50+IV-M

  • € 830,00
  • 1 Kit size L (+1000 mg EdU)
  • BCK-EdU555FC50+IV-L

  • € 1.070,00
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  • EdU in vivo cell proliferation Kit provides a superior alternative to BrdU and [3H]-thymidine assays for detection of cell proliferation in animals.
    In principal, EdU (5-ethynyl-2’-deoxyuridine), a thymidine analog, is administered to animals. EdU is incorporated into DNA during DNA synthesis and has a clickable functionality.

    Detection of cell division in tissues can be performed using microscopy, flow cytometry or in high throughput screening methods. The in vivo kits are thus designed in a sense that we provide additional EdU for the administering step. The three variants of baseclick EdU in vivo kits are:

    1. Imaging kit (BCK-EdU-IM100+IV)
    2. Flow cytometry kit (BCK-EdU-FC50+IV)
    3. High throughput screening – HTS kit (BCK-EdU-HTS2+IV)

     

    LITERATURE

    An efficient protocol for in vivo labeling of proliferating epithelial cells, C. Michel et al., 2018, J. Immunol. Methods, Vol. 457, p. 82-86.

    https://doi.org/10.1016/j.jim.2018.03.015

    Cisplatin-induced DNA double-strand breaks promote meiotic chromosome synapsis in PRDM9-controlled mouse hybrid sterility, L. Wang et al., 2018, eLife, Vol. 7, p. e42511.

    https://doi.org/10.7554/eLife.42511

    FXR Regulates Intestinal Cancer Stem Cell Proliferation, T. Fu et al., 2019, Cell, Vol. 176, p. 1098-1112.

    https://doi.org/10.1016/j.cell.2019.01.036

    FAQ

    • What type of cells can incorporate EdU?

      The EdU cell proliferation assay has been applied to many different cell types and organisms from prokaryotic to eukaryotic. Cell lines such as E. coli, 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 pathway that can phosphorylate EdU to the corresponding triphosphate, which is then accepted by the host DNA polymerase for incorporation into DNA during replication.

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

      EdU is incorporated into living cells, but the detection reaction must be performed on fixed and permeabilized samples.

    • 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 should not be used with dyes of 488 nm wavelengths.

    • When can I safely interrupt the experiment?

      It is possible to safely interrupt the protocol after the fixation step. Thereto, remove the fixation solution and wash as suggested by the user manual, then the cells can be stored in buffer at 4° C. Alternatively, the experiment can also be safely interrupted after permeabilization, again as described above.
      Please note: It is important to proceed with the experiment if the click cocktail for the detection of the EdU has been prepared already.

    • 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 be aware of the dye used for EdU detection. Check also the user manual for more information.

    • How to determine the EdU incubation time?

      The EdU incubation time depends on the cell type or organism, the applied EdU concentration and the experimental design. For a start it is advisable to refer to a literature 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. For longer incubation (> 1 day) the concentration should be decreased to 1-5 µM.

    • Shelf Life

      12 months unopened after receipt

    • Storage Conditions

      2-8 °C

    • Physical State

      kit system made of different components

    • CAS Number

      n.a.

    • Excitation (max)

      546 nm

    • Emission (max)

      579 nm

    • Ɛ (max)

      91.000 cm-1M-1

    • Preparation/Handling

      please see user manual of the kit

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