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EdU Click FC-488

From baseclick
ROTI®Kit for Flow Cytometry
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Storage temp. +4 °C
Transport temp. cooled
ADR 9 III
WGK 3
UN-Nr. 3077

Kit for detection of cell proliferation through Flow Cytometry. Marker: 6-FAM-Azid Abs. = 496 nm, λEm.= 516 nm)
For 50 assays.

Components

CHF502.60/Pack Qty. 

excl. VAT. | 1 kit per Pack Qty.

Art. No. 7779.1

Available at short notice
Free of shipping costs from 200 CHF
from 6 Pack Qty. CHF477.47/Pack Qty.
from 24 Pack Qty. CHF452.34/Pack Qty.
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In the general information or Product details, we have compiled extensive descriptions and downloads for each product.
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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 Flow Cytometry allow the measurement of DNA synthesis in flow cytometry. 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. The protocol includes only few steps and decreases the total amount of time.

Flowcytometry_plot
Figure: HeLa cells, treated without (A) and with (B) 10µM EdU for 2 h. Click reaction was performed using 6-FAM-Azide.
A: negative control, cells without EdU incorporation.
B: non-proliferating cells without EdU incorporation (blue coloured left peak) and proliferating cells (S-phase) having incorporated EdU and are labeled with 6-FAM Azide (green coloured right peak).
Density blots of PI (propidium iodide) stained samples without (C) and with (D) EdU incubation and followed click reaction. The y-axis presents the FL1-Fluorescence intensity and the x-axis the content of DNA measured with FL3-area. Cell cycle phases are indicated as G1, S and G2/M phase.

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



References:

1 N. Dinh Van et al., Modulation of HCV reinfection after orthotopic liver transplantation by fibroblast growth factor-2 and other non-interferon mediators. Gut 2016, 65, 1015-1023.

2 L. Pisarsky et al., Targeting Metabolic Symbiosis to Overcome Resistance to Anti-angiogenic Therapy. Cell Reports 2016, 15, 1-14.


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Technical Information
λ 488  nm
Markers 6-FAM  
EdU Click FC-488
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Downloads

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.

    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 Flow Cytometry (FC)

    The ROTI®Kits for Flow Cytometry by Carl ROTH use click chemistry to detect cell proliferation by means of a large number of fluorescent stain read-outs. In the detection procedure both, the total number of steps and the processing time are significantly reduced. The simple click chemistry detection procedure is complete within 30 minutes.

    References:1 N. Dinh Van et al., Modulation of HCV reinfection after orthotopic liver transplantation by fibroblast growth factor-2 and other non-interferon mediators. Gut 2016, 65, 1015-1023.2 L. Pisarsky et al., Targeting Metabolic Symbiosis to Overcome Resistance to Anti-angiogenic Therapy. Cell Reports 2016, 15, 1-14.

    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

    EdU10 mg
    6-FAM-Azide (10 mM in DMSO)130 µl
    DMSO5 ml
    Fixative Solution5 ml
    Saponin Reagent50 ml
    Catalyst Solution2 ml
    Buffer Additive400 mg

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