Janelia Fluor® 549, SE
Fluorescent dye; supplied as an NHS ester for coupling to primary amine groups. Compatible with self-labeling tag systems, e.g. HaloTag® and SNAP-tag®. Suitable for confocal fluorescent imaging and super resolution microscopy (SRM) techniques, such as dSTORM (live and fixed cells). Cell permeable. Excitation maximum = 549 nm; emission maximum = 571 nm; Quantum yield = 0.88; Extinction coefficient = 101,000 M-1cm-1; A280 correction factor is 0.169.
HaloTag is a trademark of Promega Corporation, and SNAP-tag is a trademark of New England BioLabs, Inc.
Sold under license from the Howard Hughes Medical Institute, Janelia Research Campus
Application of Janelia Fluor® Dyes in Cardiac Tissue
Application of Janelia Fluor® Dyes in Cardiac Tissue - Left image - Widefield fluorescence image of muscle tissue from an adult pig heart, displaying the distribution of collagen VI in the interstitial space between heart muscle cells. This sample was labelled with a primary antibody against collagen VI and a secondary antibody conjugated to JF549 (Cat.No. 6147). Scale bar: 50 μm. Middle image - An adult pig heart tissue section, 10 μm thick, labelled with antibodies against SERCA2ATPase, exhibits the intricate structure of the sarcoplasmic reticulum. A super-resolution image (top panel), obtained by exploiting spontaneous photo-switching of JF549 (dSTORM), shows far superior detail than the diffraction limited widefield image (bottom). Scale bar: 200 nm. Right image - A single isolated Wistar rat cardiomyocyte stained against α-actinin, displaying its periodic structure localised at the ends of sarcomeres (Z-discs). Widefield fluorescence of JF549 (left panel), prior to illuminating with a 561 nm laser to induce photoswitching and produce a dSTORM super-resolution image (right). Scale bar: 1 μm.
All images kindly provided by Prof. Christian Soeller, University of Exeter; acquired by Alex Clowsley and Anna Meletiou.
|Storage||Store at -20°C|
The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.
All Tocris products are intended for laboratory research use only.
|Solvent||Max Conc. mg/mL||Max Conc. mM|
Preparing Stock Solutions
The following data is based on the product molecular weight 551.56. Batch specific molecular weights may vary from batch to batch due to solvent of hydration, which will affect the solvent volumes required to prepare stock solutions.
|Concentration / Solvent Volume / Mass||1 mg||5 mg||10 mg|
|1 mM||1.81 mL||9.07 mL||18.13 mL|
|5 mM||0.36 mL||1.81 mL||3.63 mL|
|10 mM||0.18 mL||0.91 mL||1.81 mL|
|50 mM||0.04 mL||0.18 mL||0.36 mL|
References are publications that support the biological activity of the product.
Grimm et al (2015) A general method to improve fluorophores for live-cell and single-molecule microscopy. Nat.Methods 12 244 PMID: 25599551
Deng et al (2015) CASFISH: CRISPR/Cas9-mediated in situ labeling of genomic loci in fixed cells. Proc.Natl.Acad.Sci.USA. 112 11870 PMID: 26324940
Knight et al (2015) Dynamics of CRISPR-Cas9 genome interrogation in living cells. Science 350 823 PMID: 26564855
Legant et al (2016) High-density three-dimensional localization microscopy across large volumes. Nat.Methods 13 359 PMID: 26950745
If you know of a relevant reference for Janelia Fluor® 549, SE, please let us know.
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Protocols for Janelia Fluor® 549, SE
The following protocol features additional information for the use of Janelia Fluor® 549, SE (Cat. No. 6147).
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