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EdU Click-555

From baseclick
ROTI®Kit for Imaging
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Related products
Pack Qty.
Pack.
Storage temp. +4 °C
Transport temp. cooled
ADR 9 III
WGK 3
UN-Nr. 3077

For fast and simple detection of cell proliferation through fluorescence microscopy. Marker: 5-TAMRA-PEG3-AzidAbs. = 546 nm, λEm.= 579 nm)
For 100 Assays.

Components
€390.80 /Pack Qty.

€293.05/Pack Qty.  Campaign price!

excl. VAT. | 1 kit per Pack Qty.

Art. No. 7775.1

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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 detection of cell proliferation is of utmost importance for monitoring of cell vitality, determining genotoxicity or evaluating anticancer drugs. Carl ROTH's cell proliferation assays based on EdU (5-ethynyl-2'-deoxyuridine) provide a superior alternative to BrdU (bromodeoxyuridine) assays.
Carl ROTH's EdU Click assays are not antibody based and therefore do not require DNA denaturation for detection of the incorporated nucleoside. Instead, the EdU Click assay utilizes click chemistry for detection in a variety of dye fluorescent readouts. Furthermore, the streamlined detection protocol both reduces the total number of steps and significantly decreases the total amount of time.

Proliferierendes_Gewebe
Figure: Proliferating tissue of barley meristem labelled via EdU Click-555 kit (Art. No. 7775.1). Dividing cells incorporated EdU and were labelled via click reaction with TAMRA (pink stained cells). Nuclei were counterstained with DAPI (blue). Magnification: 100x; Filterset: Sp. Aqua HC mFISH; Sp. Green HC mFISH.

The number at the end of the product name specifies the wavelength at which the included fluorescent marker is optimally excited.



References:


S. Vidak et al., Proliferation of progeria cells is enhanced by lamina-associated polypeptide 2α (LAP2α) through expression of extracellular matrix proteins, Genes & Development 2015, 29, 2022-2036.


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-555 ROTI®Kit for Imaging

S. Barberán, S. Fraguas and F. Cebrià, The EGFR signaling pathway controls gut progenitor differentiation during planarian regeneration and homeostasis. Development 2016, 143, 2089-2102.

M. Hennenberg et al., Cooperative effects of EGF, FGF, and FGF-ß1 in prostate stromal cells are different from responses to single growth factors. Life Sciences 2015, 123, 18-24.

D. Herrmann et al., Honokiol, a constituent of Magnolia species, inhibits adrenergic contraction of human prostate strips and induces stromal cell death. Prostate Int 2014, 2, 3, 140-146.



Technical Information
λ 555  nm
Markers 5-TAMRA  
EdU Click-555
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Downloads

General information

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.

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™

    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)

  • ROTI®Kits for Imaging

    The detection of cell proliferation is of utmost importance for monitoring of cell vitality, determining genotoxicity or evaluating anticancer drugs. Carl ROTH’s cell proliferation assays based on EdU (5-ethynyl-2'-deoxyuridine) provide a superior alternative to BrdU (bromodeoxyuridine) assays. Carl ROTH’s EdU Click assays are not antibody based and therefore do not require DNA denaturation for detection of the incorporated nucleoside. Instead, the EdU Click assay utilizes click chemistry for detection in a variety of dye fluorescent readouts. Furthermore, the streamlined detection protocol both reduces the total number of steps and significantly decreases the total amount of time.

    References:
    1 S. Vidak et al., Proliferation of progeria cells is enhanced by lamina-associated polypeptide 2α (LAP2α) through expression of extracellular matrix proteins, Genes & Development 2015, 29, 2022-2036.

    EdU Click-488:

    2 V. Fock et al., Trophoblast subtype-specific EGFR/ ERBB4 expression correlates with cell cycle progression and hyperplasia in complete hydatidiform moles. Human Reproduction 2015, 30, 4, 789-799.
    3 K. Plessl et al., Expression pattern and function of Notch2 in different subtypes of first trimester cytotrophoblast. Placenta 2015, 36, 365-371.
    4 G. Meinhardt et al., Wingless ligand 5a is a critical regulator of placental growth and survival. Scientific reports 2016, 6, 28127.
    5 V. Ziegler, A. Albers, G. Fritz, Lovastatin protects keratinocytes from DNA damage-related pro-apoptotic stress responses stimulated by anticancer therapeutics. Biochimica et Biophysica Acta 2016, 1863, 1082-1092.

    EdU Click-555:

    6 S. Barberán, S. Fraguas and F. Cebrià, The EGFR signaling pathway controls gut progenitor differentiation during planarian regeneration and homeostasis. Development 2016, 143, 2089-2102.
    7 M. Hennenberg et al., Cooperative effects of EGF, FGF, and FGF-ß1 in prostate stromal cells are different from responses to single growth factors. Life Sciences 2015, 123, 18-24.
    8 D. Herrmann et al., Honokiol, a constituent of Magnolia species, inhibits adrenergic contraction of human prostate strips and induces stromal cell death. Prostate Int 2014, 2, 3, 140-146.

    Analysis certificates

    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

    EdU5 mg  
    5-TAMRA-PEG3-Azide (10 mM in DMSO)130 µl  
    Buffer Additive4 x 200 mg  
    DMSO2 x 2 ml  
    Catalyst Solution2 x 2 ml  
    Reaction Buffer4 x 2 ml  
    Store at. –20 °C (EdU, 5-TAMRA-PEG3-Azide, Buffer Additive); +4 °C (DMSO, Catalyst Solution, Reaction Buffer).

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