Janelia Fluor® 549, NHS ester
Chemical Name: 3,6-Di-1-azetidinyl-9-[2-carboxy-5-[[(2,5-dioxo-1-pyrrolidinyl)oxy]carbonyl]phenyl]xanthylium, inner salt
Biological Activity
Key Information: Janelia Fluor® 549, NHS ester is a yellow fluorescent dye; supplied with an NHS ester reactive group for the labeling of primary amines. Suitable for live cell imaging.Application: Suitable for flow cytometry, confocal microscopy, super resolution microscopy (SRM) including dSTORM and STED. Janelia Fluor® 549, NHS ester is cell permeable.
Properties and Photophysical Data: NHS ester can be converted to relevant substrate for use in self-labeling tag systems, e.g. HaloTag® and SNAP-tag®. Excitation and emission maxima (λ) are 549 nm and 571 nm, respectively; quantum yield = 0.88; extinction coefficient = 101,000 M-1cm-1; A280 correction factor = 0.169; lactone-zwitterion equilibrium constant (KL-Z) = 3.5.
Please see the protocol for further information on protein/antibody labeling and conjugation.
We also offer Janelia Fluor® conjugated antibodies and custom conjugation services.
HaloTag is a trademark of Promega Corporation, and SNAP-tag is a trademark of New England BioLabs, Inc.
Janelia Fluor is a registered trademark of Howard Hughes Medical Institute.
This product has been recently renamed. The previous name for this product was Janelia Fluor® 549, SE.
Scientific Data
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Application of Janelia Fluor® 549, NHS ester in Cardiac Tissue. An adult pig heart tissue section, 10 μm thick, labelled with abs against SERCA2ATPase, exhibits the intricate structure of the sarcoplasmic reticulum. A super-resolution image (top), obtained by exploiting spontaneous photo-switching of JF549 (dSTORM), shows superior detail to the diffraction limited widefield image (bottom). Scale: 200 nm. Image kindly provided by Prof. Christian Soeller, University of Exeter; acquired by Alex Clowsley and Anna Meletiou.
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Application of Janelia Fluor® 549, NHS ester in Cardiac Tissue. A single isolated Wistar rat cardiomyocyte stained against alpha -actinin, displaying its periodic structure localised at the ends of sarcomeres (Z-discs). Widefield fluorescence of JF549 (left), prior to illuminating with a 561 nm laser to induce photoswitching and produce a dSTORM super-resolution image (right). Scale: 1 μm. Image kindly provided by Prof. Christian Soeller, University of Exeter; acquired by Alex Clowsley and Anna Meletiou.
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Application of Janelia Fluor® Dyes in Cardiac Tissue. Widefield fluorescence image of muscle tissue from an adult pig heart, displaying the distribution of collagen VI in the interstitial space between muscle cells. The sample was labelled with a primary ab against collagen VI and a secondary ab conjugated to JF549, NHS ester. Scale: 50 μm. Image kindly provided by Prof. Christian Soeller, University of Exeter; acquired by Alex Clowsley and Anna Meletiou.
Optical Data for Janelia Fluor® 549, NHS ester
| Emission Color | Yellow |
|---|---|
| Brightness | |
| λabs | 549 nm |
| λem | 571 nm |
| Extinction Coefficient (ε) | 101,000 M-1cm-1 |
| Quantum Yield (φ) | 0.88 |
| Closest Laser line | 532 nm |
| Reactive Group | NHS ester |
| Reactivity | Primary amines |
| Cell Permeable | Yes |
| Correction Factor 280 | 0.169 |
| Application | Confocal Microscopy, Flow Cytometry, Super Resolution Microscopy, dSTORM, STED |
| Alternative For | Alexa Fluor® 546, Alexa Fluor® 555, BDY TMR-X, Atto 550, CF 555, TAMRA, Cyanine 3 |
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company.
Plan Your Experiments
Use our spectra viewer to interactively plan your experiments, assessing multiplexing options. View the excitation and emission spectra for our fluorescent dye range and other commonly used dyes.
Spectral Viewer
Technical Data
The technical data provided above is for guidance only.
For batch specific data refer to the Certificate of Analysis.
Tocris products are intended for laboratory research use only, unless stated otherwise.
Additional Information
Background References
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A general method to improve fluorophores for live-cell and single-molecule microscopy.
Grimm et al.
Nat.Methods, 2015;12:244 -
High-density three-dimensional localization microscopy across large volumes.
Legant et al.
Nat.Methods, 2016;13:359 -
CASFISH: CRISPR/Cas9-mediated in situ labeling of genomic loci in fixed cells.
Deng et al.
Proc.Natl.Acad.Sci.USA., 2015;112:11870 -
Dynamics of CRISPR-Cas9 genome interrogation in living cells.
Knight et al.
Science, 2015;350:823 -
Rational design of fluorogenic and spontaneously blinking labels for super-resolution imaging.
Zheng et al.
ACS Cent.Sci., 2019;5:1602
Product Datasheets
Citations for Janelia Fluor® 549, NHS ester
The citations listed below are publications that use Tocris products. Selected citations for Janelia Fluor® 549, NHS ester include:
8 Citations: Showing 1 - 8
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Improved Methods for Single-Molecule Fluorescence In Situ Hybridization and Immunofluorescence in Caenorhabditis elegans Embryos.
Authors: Dylan M Et al.
Curr Protoc 2021;1:e299
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Precision of tissue patterning is controlled by dynamical properties of gene regulatory networks.
Authors: James Et al.
Development 2021;148
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Internalization of the Membrane Attack Complex Triggers NLRP3 Inflammasome Activation and IL-1β Secretion in Human Macrophages.
Authors: Daniel M Et al.
Front Immunol 2021;12:720655
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SNX19 restricts endolysosome motility through contacts with the endoplasmic reticulum.
Authors: Juan S Et al.
Nat Commun 2021;12:4552
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mRNA localization is linked to translation regulation in the Caenorhabditis elegans germ lineage.
Authors: Florian Et al.
Development 2020;147
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mRNA localization is linked to translation regulation in the Caenorhabditis elegans germ lineage.
Authors: Erin Osborne Et al.
Development 2020;
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Visualizing Transcription Factor Binding on Mitotic Chromosomes Using Single-Molecule Live-Cell Imaging.
Authors: Sheila S Et al.
Methods Mol Biol 2019;2038:239-250
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Detection and quantification of RNA decay intermediates using XRN1-resistant reporter transcripts.
Authors: Franka Et al.
Nat Protoc 2019;14:1603-1633
FAQs
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