Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *High Sensitivity*
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Telephone | 1-800-990-8053 |
Fax | 1-800-609-2943 |
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Excitation (nm) | 502 |
Emission (nm) | 522 |
H-phrase | H303, H313, H340 |
Hazard symbol | T |
Intended use | Research Use Only (RUO) |
R-phrase | R20, R21, R68 |
UNSPSC | 41116134 |
Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *Optimized for Microplate Readers* |
Helixyte™ Green Fluorimetric dsDNA Quantitation Kit *Optimized for Broad Dynamic Range* |
Overview | SDSProtocol |
Excitation (nm) 502 | Emission (nm) 522 |
Platform
Spectrofluorometer
Excitation | 490 nm |
Emission | 525 nm |
Cutoff | 515 nm |
Components
Example protocol
AT A GLANCE
Protocol summary
- Add 1mL dsDNA standards or test samples to each cuvette
- Add 1mL Helixyte Green™ working solution
- Incubate at RT for 5-10 minutes
- Monitor the fluorescence at Ex/Em=490/525 nm
Important notes
The following protocol is an example for quantifying dsDNA with Helixyte Green™. Allow all the components to warm to room temperature before opening. No data is available addressing the mutagenicity or toxicity of Helixyte Green™dsDNA stain. Because this reagent binds to nucleic acids, it should be treated as a potential mutagen and handled with appropriate care. The DMSO stock solution should be handled with particular caution as DMSO is known to facilitate the entry of organic molecules into tissues.
PREPARATION OF STOCK SOLUTION
1. Assay Buffer (1X)
Prepare a 1X Assay buffer by diluting the concentrated buffer 20-fold with sterile, distilled, DNase-free water.
PREPARATION OF STANDARD SOLUTION
For convenience, use the Serial Dilution Planner: https://www.aatbio.com/tools/serial-dilution/17651
For high range standard curve:
Add 30 µL of 100 µg/mL dsDNA stock solution (Component C) to 1.47 mL of 1X Assay buffer to have 2000 ng/mL dsDNA solution, and then perform 1:2 and 1:10 serial dilutions to get 1000, 100, 10, 1 and 0 ng/mL.
For low range standard curve:
Add 40 µL of 2 µg/mL dsDNA stock solution to 1.56 mL of 1X Assay buffer to have 50 ng/mL dsDNA solution, and then perform 1:2 and 1:10 serial dilutions to get 25, 2.5, 0.25, 0.025 and 0 ng/mL.
PREPARATION OF WORKING SOLUTION
Prepare Helixyte Green™ working solution by making a 200-fold dilution of the concentrated DMSO solution in 1X assay buffer. For example, to prepare enough working solution to assay 10 samples in a 2 mL final volume, add 50 μL of Helixyte Green™ (Component A) into 10 mL of Assay Buffer (Component B). Protect the working solution from light by covering it with foil or placing it in the dark. Note: We recommend preparing this solution in a plastic container rather than glass, as the dye may adsorb to glass surfaces. For best results, this solution should be used within a few hours of its preparation.
SAMPLE EXPERIMENTAL PROTOCOL
- Add 1 mL of Helixyte Green™ working solution to each cuvette containing 1 mL of the dsDNA standard, blank control, and test samples to make the total dsDNA assay volume of 2 mL/cuvette.
- Incubate the reaction at room temperature for 5 to 10 minutes, protected from light.
- Monitor the fluorescence increase with a spectroflurometer at Ex/Em = 490/525 nm. Note: To minimize photobleaching effects, keep the time for fluorescence measurement constant for all samples.
Product Family
Name | Excitation (nm) | Emission (nm) |
Helixyte™ Green Fluorimetric ssDNA Quantitation Kit *Optimized for Microplate Readers* | 498 | 519 |
Images
Citations
Authors: Barsa, Chloe and Perrin, Julian and David, Claudine and Mourier, Arnaud and Rojo, Manuel
Journal: bioRxiv (2024): 2024--03
References
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Journal: Parasitol Res (2010): 933
Authors: Dragan AI, Bishop ES, Casas-Finet JR, Strouse RJ, Schenerman MA, Geddes CD.
Journal: J Immunol Methods (2010): 95
Authors: Moreno LA, Cox KL.
Journal: J Vis Exp. (2010)
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Journal: J Agric Food Chem (2009): 7221
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Journal: J Pharm Biomed Anal (2009): 997
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Journal: Langmuir (2009): 43
Application notes
Fluorescent Oligonucleotide Labeling Reagents
Monitoring of Mitochondrial Membrane Potential Changes in Live Cells Using JC-10
Selective Analysis of RNA in Live and Fixed Cells with StrandBrite RNA Green
Cell Loading Protocol For Fluorescent pH Indicator, BCECF-AM
FAQ
I ordered your phalloidin-amine (Cat #5302) so I can conjugate it to my oligo. Do you have a recommended protocol I can use?
What dye works best for staining and tracking lysosomes in live cells for several hours?
How can I lyse my cells without lysing the nuclear membrane?
Do you have any dual-fluorescence nucleic acid stains that interact with both DNA and RNA?