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EdU Click HTS 20-488

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ROTI®Kit for High Throughput Screening
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Related products
Pack Qty.
Pack.
Storage temp. +4 °C
Transport temp. cooled
ADR 9 III
WGK 3
UN-Nr. 3082

For fast and specific detection of cell proliferation in High Throughput Screening. Marker: 6-FAM-Azid Abs. = 496 nm, λEm.= 516 nm)
For 2000 reactions or 20 x 96 well plates.

Components

€1,059.00/Pack Qty. 

excl. VAT. | 1 kit per Pack Qty.

Art. No. 7788.1

Available from stock Karlsruhe
Delivery fast, simple and reliable!
from 6 Pack Qty. €1,006.05/Pack Qty.
from 24 Pack Qty. €953.10/Pack Qty.
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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.

EdU_HTS_KIT_well_C9_und_G9
Figure: Fluorescence imaging of 2 wells: Well C9 incubated for 6 h with EdU; Well G9: Incubation without EdU.

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.

Proliferation_HeLa
Figure: HeLa cells incubated without and with 10 µM EdU for 2, 4 or 6 h in 96 well plate. EdU incorporation was detected over time and for different cell numbers using a fluorescence plate reader. Control: Without EdU no fluorescence was detected after 6 h incubation time.

Important information on refrigerated transports

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 20-488 ROTI®Kit for High Throughput Screening

Proliferation_HeLa
Figure: HeLa cells incubated without and with 10 µM EdU for 2, 4 or 6 h in 96 well plate. EdU incorporation was detected over time and for different cell numbers using a fluorescence plate reader. Control: Without EdU no fluorescence was detected after 6 h incubation time.

EdU_HTS_KIT_well_C9_und_G9
Figure: Fluorescence imaging of 2 wells: Well C9 incubated for 6 h with EdU; Well G9: Incubation without EdU.


Technical Information
λ 488 nm
Markers 6-FAM 
EdU Click HTS 20-488
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7788.1 1 kit cardboard €1,059.00
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General information

Tip:

The number at the end of the product name specifies the excitation wavelength of the included fluorescent marker. Suitable excitation source are:

  • 488 nm (blue-green argon laser 488 nm, blue diode laser 473 nm)
  • 555 nm (green Nd:YAG laser 532 nm, green helium-neon-laser 543 nm)
  • 594 nm (yellow helium-neon-laser 594 nm, yellow diode laser 594 nm)
  • 647 nm (red krypton laser 647 nm, red helium-neon-laser 633 nm, red diode laser 635 or 650 nm)

  • Cell Proliferation Assays

    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


    Code in product name Markers Absorption (nm) Emission (nm) Filter configuration
    -488 6-FAM azide 496 516 FITC
    -555 5-TAMRA azide 546 579 Cy3™
    -594 5/6 Sulforhodamine 584 603 Texas Red®
    -647 Eterneon red (analogous to cyanine 5 azide) 643 662 Cy5T™

    Click Chemistry

    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.


    ROTI®Kits for High Throughput Screening (HTS)

    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.


    ROTI®Kits for High Throughput Screening (HTS)

    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

    You can search for and download your certificate of analysis for the selected product here. Please provide your batch number.
    The following analysis certificates have been found:

    Components

    20x EdU20 ml  
    6-FAM-Azide (10 mM in DMSO)9 x 130 µl  
    Reaction Buffer4 x 50 ml  
    Catalyst Solution5 ml  
    Buffer Additive2 x 1 g  
    Rinse Buffer58 ml  
    Store at +4 °C (20x EdU, 6-FAM-Azide, Catalyst Solution); +15 to +25 °C (Reaction Buffer, Buffer Additive, Rinse Buffer).

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