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Janelia Fluor® 646, SE
Fluorogenic fluorescent dye; supplied as an NHS ester for coupling to primary amine groups. NHS ester can be converted to relevant substrate for use in self-labeling tag systems, e.g. HaloTag® and SNAP-tag®. Suitable for confocal fluorescent imaging, super resolution microscopy (SRM) techniques such as dSTORM (live and fixed cells) and STED imaging. Can be multiplexed for two colour imaging with Janelia Fluor® 549 SE (Cat. No. 6147). Also suitable for flow cytometry. Cell permeable. Excitation maximum = 646 nm; emission maximum = 664 nm. Quantum yield = 0.54, Max. extinction coefficient = 152,000 M-1cm-1 (measured in ethanol plus 0.1% TFA); A280 correction factor is 0.19.
We recommend that stock solutions of this dye are prepared in anhydrous DMF.
To measure the absorbance spectrum of this dye we recommend the following solvent: ethanol or trifluoroethanol plus 0.1% TFA.
We also offer Janelia Fluor® conjugated antibodies and custom conjugation services with our sister company Novus Biologicals.
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
Super-resolution image (dSTORM) of muscle tissue from the left ventricle of an adult pig heart. JF646 (Cat.No. 6148) was used to label collagen VI, which is present on the cell plasma membrane, and imaged with a 642 nm excitation laser. A clearer structure is achieved in comparison with the widefield image (top right). Scale bar 2 μm.
Image kindly provided by Prof. Christian Soeller, University of Exeter; acquired by Alex Clowsley and Anna Meletiou.
Optical Data for Janelia Fluor® 646, SE
|Extinction Coefficient (ε)||152,000 M-1cm-1|
|Quantum Yield (φ)||0.54|
|Closest Laser line||640 nm|
|Reactive Group||NHS ester|
|Correction Factor 280||0.19|
|Application||Confocal imaging, dSTORM, STED, Flow Cytometry|
|Alternative For||Alexa Fluor® 647, Atto 647, Atto 647N, CF640R, CF647, Cyanine 5|
Alexa Fluor® is a registered trademark of Molecular Probes, Inc, a Thermo Fisher Scientific Company.
|Storage||Store at -20°C|
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.
|Solvent||Max Conc. mg/mL||Max Conc. mM|
Preparing Stock Solutions
The following data is based on the product molecular weight 593.71. Batch specific molecular weights may vary from batch to batch due to the degree of hydration, which will affect the solvent volumes required to prepare stock solutions.
|Concentration / Solvent Volume / Mass||1 mg||5 mg||10 mg|
|0.5 mM||3.37 mL||16.84 mL||33.69 mL|
|2.5 mM||0.67 mL||3.37 mL||6.74 mL|
|5 mM||0.34 mL||1.68 mL||3.37 mL|
|25 mM||0.07 mL||0.34 mL||0.67 mL|
References are publications that support the biological activity of the product.
Schmidt et al (2016) Live cell imaging reveals the dynamics of telomerase recruitment to telomeres. Cell 166 1188 PMID: 27523609
Li et al (2016) Real-time imaging of Huntingtin aggregates diverting target search and gene transcription. Elife 5 e17056 PMID: 27484239
Legant et al (2016) High-density three-dimensional localization microscopy across large volumes. Nat.Methods 13 359 PMID: 26950745
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
Ticau et al (2015) Single-molecule studies of origin licensing reveal mechanisms ensuring bidirectional helicase loading. Cell 161 513 PMID: 25892223
Hong et al (2009) Phosphorylation of the RNA polymerase II C-terminal domain by TFIIH kinase is not essential for transcription of Saccharomyces cerevisiae genome. Proc.Natl.Acad.Sci.U.S.A. 106 14276 PMID: 19666497
If you know of a relevant reference for Janelia Fluor® 646, SE, please let us know.
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Citations for Janelia Fluor® 646, SE
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Reviews for Janelia Fluor® 646, SE
Average Rating: 4 (Based on 1 Review.)
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I used this dye for labeling bacterial proteins in vitro. The labeling method is pretty straight-forward as for any NHS-protein coupling. The dye is highly photostable as determined by single protein tracking on lipid bilayers (image of single proteins attached, scale bar is 5 micron). The only reason I give the product 4 stars instead of 5 is the packaging. The dye is shipped as a solid in a glass bottle with a flexible rubber cap. Most of the dye gets stuck inside and on the surface of this cap; thus not delivering a full value for money. I will highly recommend the manufacturers to use bottles with plastic caps so that the dye can be recovered a 100%
Protocols for Janelia Fluor® 646, SE
The following protocol features additional information for the use of Janelia Fluor® 646, SE (Cat. No. 6148).
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