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    products/reagents/draq5-ab108410.pdf

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Epigenetics and Nuclear Signaling DNA / RNA DNA / Nucleotides
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DRAQ5™ (ab108410)

  • Datasheet
  • SDS
Reviews (5)Q&A (32)References (53)

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Immunocytochemistry/ Immunofluorescence - DRAQ5™ (ab108410)
  • Immunocytochemistry/ Immunofluorescence - DRAQ5™ (ab108410)
  • Immunocytochemistry/ Immunofluorescence - DRAQ5™ (ab108410)

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Overview

  • Product name

    DRAQ5™
  • Tested applications

    Suitable for: FM, Flow Cyt, ICC/IFmore details
  • General notes

    DRAQ5™ is a cell permeable far-red fluorescent DNA dye that can be used in fixed or non-fixed/ live cells in combination with common labels such as GFP or FITC.

    As with any cell­-permeant DNA intercalating probe, DRAQ5 may inhibit cell division in long-term assays and should be tested for any effect.

    DRAQ5 staining can be used in flow cytometry, live cell imaging and cell-based assays and the dye is highly compatible with standard protocols across many instrumentation platforms.

    The chemical name of DRAQ5 is 1, 5-bis{[2-(di-methylamino)ethyl]amino}-4, 8-dihydroxyanthracene-9, 10-dione.

    The advantages of DRAQ5 staining include
    - convenient ready-to-use aqueous solution
    - rapid uptake into living cells, providing a high level of nuclear discrimination
    - no photobleaching effect
    - can be used in most cell types, eukaryotic and prokaryotic: mammalian, bacterial, parasitic, plant, etc.
    - no compensation needed with common FITC/GFP + PE combinations in flow cytometry
    - no RNase treatment required
    - no fluorescence enhancement upon DNA binding
    - compatible with optics of benchtop flow, laser scanning cytometers and non-UV laser scanning and lamp-based confocal microscopes

    SPECTRAL PROPERTIES:

    Excitation

    • 647 nm line optimal (Exmax 646 nm)
    • 488, 514, 568 and 633 nm lines, sub-optimal
    • Two-photon excitation (1047 nm) and excitation dark (700-850 nm)

    Emission (instrument dependent):
    - 665 nm to infra-red max 681 nm / 697 nm intercalated with dsDNA)
    - minimal overlap with vis range e.g. GFP and FITC
    - Em. filters may include 695L, 715LP or 780 LP

     

     

    Concentration: 5 mM

Properties

  • Form

    Liquid
  • Storage instructions

    Store at +4°C. Do Not Freeze. Store In the Dark.
  • Concentration information loading...
  • Research areas

    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA / Nucleotides
    • Kits/ Lysates/ Other
    • Tools and Reagents
    • IHC Tools/ Reagents
    • Kits/ Lysates/ Other
    • Tools and Reagents
    • Fluorescent dyes and reagents
    • Kits/ Lysates/ Other
    • Kits
    • Cell Staining Kits
    • Fluorescent
    • Kits/ Lysates/ Other
    • Kits
    • Cell Staining Kits
    • Nuclear

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab108410 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
FM
Use at an assay dependent concentration.
Flow Cyt
1/250 - 1/1000.

For Gating of Nucleated cells = 5µM; For Cell Cycle Analysis = 20µM

ICC/IF
1/1000. For Cell-based assays, Immunofluorescence microscopy and In-Cell WB = 5µM
It is highly recommended that the concentration and labelling conditions are carefully determined by each investigator for optimal performance in the assay of interest. For more specific information about the applications, please refer to the Protocol Booklet section.
Notes
FM
Use at an assay dependent concentration.
Flow Cyt
1/250 - 1/1000.

For Gating of Nucleated cells = 5µM; For Cell Cycle Analysis = 20µM

ICC/IF
1/1000. For Cell-based assays, Immunofluorescence microscopy and In-Cell WB = 5µM
It is highly recommended that the concentration and labelling conditions are carefully determined by each investigator for optimal performance in the assay of interest. For more specific information about the applications, please refer to the Protocol Booklet section.

Images

  • Immunocytochemistry/ Immunofluorescence - DRAQ5™ (ab108410)
    Immunocytochemistry/ Immunofluorescence - DRAQ5™ (ab108410)

    HeLa cells were stained with Lamin B1 antibody - Nuclear Envelope Marker (ab16048) and alpha Tubulin antibody [DM1A] - Loading Control (ab7291). The cells were 100% methanol fixed (5 min) and then incubated in 1% BSA in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the primary antibodies (ab16048 & ab7291) at 1µg/ml overnight at 4C. The secondary antibodies were Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488), pre-adsorbed (ab96899) (green) and Goat polyclonal Secondary Antibody to Mouse IgG - H&L (DyLight® 594), pre-adsorbed (ab96881) (red) used at 1/250 dilution for 1h at room temperature. 5µM DRAQ5 was added to the secondary antibody mixture to label nuclear DNA (pseudocolor purple).

  • Immunocytochemistry/ Immunofluorescence - DRAQ5™ (ab108410)
    Immunocytochemistry/ Immunofluorescence - DRAQ5™ (ab108410)

    HeLa cells were stained with beta Catenin antibody (ab16051) and alpha Tubulin antibody [DM1A] - Loading Control (ab7291). The cells were 100% methanol fixed (5 min) and then incubated in 1% BSA in 0.1% PBS-Tween for 1h to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the primary antibodies (ab16051 & ab7921) at 1µg/ml overnight at 4C. The secondary antibodies were Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 488), pre-adsorbed (ab96899) (green) and Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (DyLight® 594), pre-adsorbed (ab96899) (red) used at 1/250 dilution for 1h at room temperature. 5µM DRAQ5 was added to the secondary antibody mixture to label nuclear DNA (pseudocolor orange).

  • Immunocytochemistry/ Immunofluorescence - DRAQ5™ (ab108410)
    Immunocytochemistry/ Immunofluorescence - DRAQ5™ (ab108410)
    DRAQ5™-stained nuclei in a adult Drosophila brain.

Protocols

  • Protocol Booklet

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (53)

Publishing research using ab108410? Please let us know so that we can cite the reference in this datasheet.

ab108410 has been referenced in 53 publications.

  • Yap EL  et al. Bidirectional perisomatic inhibitory plasticity of a Fos neuronal network. Nature 590:115-121 (2021). PubMed: 33299180
  • Zhang YW  et al. Human Plasma In-Cell Western Assays-An In vitro Predictor for In vivo Pharmacology in Oncology Drug Discovery. Curr Protoc 1:e51 (2021). PubMed: 33587334
  • SenGupta S  et al. Triple-Negative Breast Cancer Cells Recruit Neutrophils by Secreting TGF-ß and CXCR2 Ligands. Front Immunol 12:659996 (2021). PubMed: 33912188
  • Senatore E  et al. The TBC1D31/praja2 complex controls primary ciliogenesis through PKA-directed OFD1 ubiquitylation. EMBO J 40:e106503 (2021). PubMed: 33934390
  • Martínez AL  et al. Development of a novel in vitro assay to screen for neuroprotective drugs against iatrogenic neurite shortening. PLoS One 16:e0248139 (2021). PubMed: 33690613
View all Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews

DRAQ5: Cell cycle analysis with flow cytometry

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
DRAQ5 has been used with LNCaP C4-2B cell line in flow cytometry for cell cycle analysis. The cells were scraped and centrifuged for 5 min at 4C at 1.200 rpm. Washed with cold PBS and further centrifuged for 5 min at 4C at 1.200 rpm. The cells were then stained with 200 ul of staining solution (DRAQ5 1:1000 dil in PBS) for 1 h at room temperature protected from lighted. The reaction volume was finally made up to 500 ul and the cells were examined by flow cytometry. There is a distinct difference in the apoptotic induced treated cells with a high apoptoptic percentage (M1) and lower M2, M3 and M4 percentages, representing G1, S and G2-M phases, respectively.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Dimitra Kalamida

Verified customer

Submitted Aug 22 2018

DRAQ5 In cell Western As Part of an Ab Trial scheme

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
DRAQ5 has been used to normalize for cell number( C42B cell line) . DRAQ5, a DNA dye has been used to label nuclei, in 1:1000 dilution in PBS, with a total volume of 50µl/ well. The cells were incubated for 30 minutes at room temperature in dark, before scanning using appropriate excitation (648 nm) and emission (705LP) filters. Cell number titration has been performed using 60000, 30000, 15000 and 7500, respectively. The fluorescence intensity of each well was quantified using ImageJ and plotted in a graph using GraphPrism, as demonstrated in following figure .
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Dimitra Kalamida

Verified customer

Submitted Jun 05 2018

So good!

Good Good 4/5 (Ease of Use)
Abreviews
Abreviews
It is easy for staining with DAPI! so good.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Ms. Sehee Oh

Verified customer

Submitted Jan 31 2018

DRAQ5 co-staining

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Excellent
Compatible with FITC, Alexa 568, and Alexa 405 co-staining
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Dec 08 2014

DRAQ5 Abcam for In Cell Western Assay

Excellent Excellent 5/5 (Ease of Use)
Abreviews
Abreviews
abreview image
Our lab uses DRAQ5 from Abcam for the In Cell Western Assay. DRAQ5 is used to provide accurate normalization over a broad range of cell densities. Specifically, it can be used for stoichiometric staining of DNA in live or fixed cells. DRAQ5 is used in the 700nm channel to normalize well-to well variation in cell number.
Picture shows In Cell Wester Assay using DRAQ5 in PC3 and LnCAP prostate cell lines.
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Jun 25 2014

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