EdU Click HTS 4-555
All products identified as being particularly temperaturesensitive are shipped in special ice boxes with freezer packs or in dry ice.
Ready-to-use products and reagents
Transport temp. cooled
ADR 9 III
WGK 3
UN-Nr. 3082
For 400 reactions or 4 x 96 well plates.
€321.50/Pack Qty.
excl. VAT. | 1 kit per Pack Qty.
Art. No. 7791.1
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Product details
- Highly reliable
- Easy to perform
- Fast detection procedure (only 30 min)
- Mild conditions (no DNA denaturation required)
- Modular system due to click chemistry
- Compatible with various dyes and readouts
- Compatible with multiplexing
- No cytotoxicity up to 1 mM EdU
The ROTI®Kits for High Throughput Screening allow the measurement of DNA synthesis adherent cells in 96-well plate format. Similar to BrdU assays, our assays utilise a uracil derivative (in this case ethynyldeoxyuridine, EdU) which is incorporated into the replicating DNA. EdU is not detected via antibodies; instead fluorescent dyes are coupled directly via click reaction. This fast and elegant approach allows for direct labelling of replicating DNA with the desired fluorescent dye. Thus, the assay is perfectly adaptable to the present facilities. Furthermore, the click reaction is highly selective and therefore prevents any unspecific labelling. The protocol includes only few steps and decreases the total amount of time.
By using Click chemistry, the detection of the fluorescent dyes used is highly specific and very fast. The kits are available in different sizes. The size (2, 4 or 20) refers to the number of 96-well plates the kit is designed for.
The number at the end of the product name specifies the wavelength at which the included fluorescent marker is optimally excited.
All products identified as being particularly temperature-sensitive are shipped in special ice boxes with freezer packs or in dry ice.
Additional costs resulting from such, will be invoiced (further information on request).
Please note: In order to guarantee optimum product quality, the dispatch of refrigerated transport products will be carried out on Mon and Tue only outside Germany!
Slight delays in delivery are therefore possible.
EdU Click HTS 4-555 ROTI®Kit for High Throughput Screening
| λ | 555 nm |
| Markers | 5-TAMRA |
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| Order No. | Pack Qty. | Pack. | Price | Quantity | ||
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| 7791.1 | 1 kit | cardboard | €321.50 |
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General information
The number at the end of the product name specifies the excitation wavelength of the included fluorescent marker. Suitable excitation source are:
Our ROTI®Kits for EdU mediated detection of cell proliferation provide a superior alternative to time-consuming and less sensitive bromodeoxyuridine (BrdU) assays. Similar to BrdU assays, our assays utilise a uracil derivative (in this case ethynyldeoxyuridine, EdU) which is incorporated into the replicating DNA. EdU is not detected via antibodies; instead fluorescent dyes are coupled directly via click reaction. This fast and elegant approach allows for direct labelling of replicating DNA with the desired fluorescent dye. Thus, the assay is perfectly adaptable to the present facilities and to the assay situation (e.g. for double or triple detection). Furthermore, the click reaction is highly selective and therefore prevents any unspecific labelling. The protocol includes only few steps and decreases the total amount of time.
Kits available for:
• Fluorescence microscopy (ROTI®Kits for Imaging)
• Flow Cytometry (ROTI®Kits for Flow Cytometry)
• High Throughput Screening (ROTI®Kits for High Throughput Screening)
Advantages of the click chemistry based approach:
Schematic illustration of the click chemistry based EdU cell proliferation assay. A) Incubation of cells with EdU. EdU is a thymidine analogue and is incorporated in DNA during active DNA synthesis. B) After DNA synthesis, EdU was incorporated in the DNA strands. C) Detection of proliferating cells via click chemistry. This reaction is finished within 30 minutes, whereby a variety of different fluorescent dye azides can be used.
Advantages of the Click Assay:
- Highly reliable
- Easy handling
- Fast detection procedure (only 30 min.)
- Mild conditions, no DNA denaturation required
- Compatible with various dyes
- Modular system in click chemistry
- Compatible with multiplexing
The copper(I)-catalysed azide-alkyne cycloaddition (CuAAC) is the most prominent example of a group of reactions named click reactions. According to Sharpless’ definition, these reactions are characterised by high yields, mild reaction conditions, and by their tolerance of a broad range of functional groups.[1-3] Typically, the reactions require simple or no workup or purification of the product. The most important characteristic of the CuAAC reaction is its unique bioorthogonality as neither azide nor terminal alkyne functional groups are generally present in natural systems.
References:
1 H. C. Kolb, M.G. Finn, K. B. Sharpless, Angew. Chem. Int. Ed. 2001, 40, 2004-2021.
2 C.W. Tornoe, C. Christensen, M. Meldal, J. Org. Chem. 2002, 67, 3057-3064.
3 V. V. Rostovtsev, L. G. Green, V. V. Fokin, K. B. Sharpless, Angew. Chem. 2002, 114, 2708-2711; Angew. Chem. Int. Ed. 2002, 41, 2596-2599.
The ROTI®Kits for High Throughput Screening allow the measurement of DNA synthesis adherent cells in 96-well plate format. Similar to BrdU assays, our assays utilise an uracil derivative (in this case ethynyldeoxyuridine, EdU) which is incorporated into the replicating DNA. EdU is not detected via antibodies; instead fluorescent dyes are coupled directly via click reaction. This fast and elegant approach allows for direct labelling of replicating DNA with the desired fluorescent dye. Thus, the assay is perfectly adaptable to the present facilities. Furthermore, the click reaction is highly selective and therefore prevents any unspecific labelling. The protocol includes only few steps and decreases the total amount of time.
The ROTI®Kits for High Throughput Screening allow the measurement of DNA synthesis adherent cells in 96-well plate format. Similar to BrdU assays, our assays utilise an uracil derivative (in this case ethynyldeoxyuridine, EdU) which is incorporated into the replicating DNA. EdU is not detected via antibodies; instead fluorescent dyes are coupled directly via click reaction. This fast and elegant approach allows for direct labelling of replicating DNA with the desired fluorescent dye. Thus, the assay is perfectly adaptable to the present facilities. Furthermore, the click reaction is highly selective and therefore prevents any unspecific labelling. The protocol includes only few steps and decreases the total amount of time.
Certificates of Analysis
Components
| 20x EdU | 4 ml |
| 5-TAMRA-PEG3-Azide (10 mM in DMSO) | 2 x 130 µl |
| Reaction Buffer | 40 ml |
| Catalyst Solution | 1 ml |
| Buffer Additive | 400 mg |
| Rinse Buffer | 2 x 6 ml |
| Store at +4 °C (20x EdU, 5-TAMRA-PEG3-Azide, Catalyst Solution); +15 to +25 °C (Reaction Buffer, Buffer Additive, Rinse Buffer). | |