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ClickTech EdU Cell Proliferation Kit 488 (200 Assays) for HTS IV
In vivo EdU Cell Proliferation Kit for High-throughput Screening
- Catalog No.
- 1 Kit size S (+50 mg EdU)
- € 355,00
- 1 Kit size M (+500 mg EdU)
- € 660,00
- 1 Kit size L (+1000 mg EdU)
- € 900,00
Our EdU in vivo cell proliferation Kits provide a superior alternative to BrdU and [3H]-thymidine assays for detection of cell proliferation in whole animals.
In principal, EdU (5-ethynyl-2’-deoxyuridine), a thymidine analog, is fed to animals. As it is incorporated into DNA during active DNA synthesis and shows a clickable functionality, the content of new cells can be assessed after harvesting the tissue of interest. Detection can then be performed after reaction with the dye of choice, using our 3 standard fluorescent readouts: microscopic imaging, flow cytometry or HTS. The in vivo kits are thus designed in a sense that we combine extra EdU for the feeding step with one of the three baseclick EdU cell proliferation kits such as:
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)
Michel, et al, J. Immunol. Methods, 2018, 457, 82-86
Wang, et al., Elife, 2018, 7, e42511
Fu, et al., Cell, 2019, 176, 1098-1112
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).
- What type of cells can incorporate EdU?
12 months unopened after receipt
kit system made of different components
please see user manual of the kit
- Shelf Life