Janelia Fluor® 646, NHS ester
Tocris Bioscience, part of Bio-Techne | Catalog # 6148
Red dye (Reactive group: NHS ester). Application: flow cytometry, confocal microscopy, super resolution microscopy (SRM) including dSTORM & STED. Suitable for live cell imaging
Key Product Details
Description
Red dye (Reactive group: NHS ester). Application: flow cytometry, confocal microscopy, super resolution microscopy (SRM) including dSTORM & STED. Suitable for live cell imaging
Alternative Names
JF 646,NHS
Product Description
Key Information: Janelia Fluor® 646, NHS ester is a red fluorogenic 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) techniques including dSTORM (in both live and fixed cells) and STED. 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®. Can be multiplexed for two color imaging with Janelia Fluor® 549 NHS ester (Cat. No. 6147). Excitation and emission maxima (λ) are 646 nm and 664 nm, respectively; quantum yield = 0.54; extinction coefficient = 152,000 M-1cm-1 (measured in TFE plus 0.1% TFA); A280 correction factor = 0.19; lactone-zwitterion equilibrium constant (KL-Z) = 0.0012.
Please see the protocol for further information on protein/antibody labeling and conjugation.
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 custom conjugation antibody services.
HaloTag® is a registered trademark of Promega Corporation, and SNAP-tag® is a registered 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® 646, SE.
Scientific Data Images for Janelia Fluor® 646, NHS ester
Super-resolution imaging of methylated histones (H3K9me3) in a COS-7 cell line.
COS-7 cells were fixed and stained with rabbit anti-histone H3K9me3 (Active Motif, Catalog # 39062, 1:1000 dilution) followed by Janelia Fluor® 646 labelled anti-rabbit IgG (H+L) secondary antibody (Novus, Catalog # NBP1-75335, 1:2000 dilution). Super-resolution images were acquired with the Sysmex HM-1000. Images were acquired as 20000 frame stack files with 30 msec. exposure time and image was constructed using HM-1000 software or ThunderSTORM.
Application of Janelia Fluor® 646, NHS ester in Cardiac Tissue.
Super-resolution image (dSTORM) of muscle tissue from the left ventricle of an adult pig heart. Janelia Fluor 646, NHS ester 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: 2 μm.Image kindly provided by Prof. Christian Soeller, University of Exeter; acquired by Alex Clowsley and Anna Meletiou.Licensing Information
Sold under license from the Howard Hughes Medical Institute, Janelia Research CampusOptical Data
| Emission Color | Red |
| Brightness | |
| λabs | 646 nm |
| λem | 664 nm |
| Extinction Coefficient (ε) | 152,000 M-1cm-1 |
| Quantum Yield (φ) | 0.54 |
| Closest Laser line | 640 nm |
| Reactive Group | NHS ester |
| Reactivity | Primary amines |
| Cell Permeable | Yes |
| Correction Factor 280 | 0.19 |
| Application | Confocal Microscopy, dSTORM, STED, Flow Cytometry |
| Alternative For | Alexa Fluor® 647, Atto 647, Atto 647N, CF640R, CF647, Cyanine 5 |
Trademarks and registered trademarks are the property of their respective owners.
Spectra Viewer
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Product Specifications
Molecular Weight
593.71
Formula
C33H31N3O6Si
Storage
Store at -20°C
Purity
≥95% (HPLC)
Chemical Name
1-[7-(1-Azetidinyl)-10-[2-carboxy-5-[[(2,5-dioxo-1-pyrrolidinyl)oxy]carbonyl]phenyl]-9,9-dimethyl-9-silaanthracen-2(9H)-ylidene]azetidinium, inner salt
CAS Number
1811539-59-9
PubChem ID
124201858
InChI Key
ODBCGJUBBYOJOS-UHFFFAOYSA-N
SMILES
O=C(ON1C(CCC1=O)=O)C2=CC=C(C([O-])=O)C(C(C3=CC=C(N4CCC4)C=C3[Si]5(C)C)=C(C=C/6)C5=CC6=[N+]7CCC\7)=C2
The technical data provided above is for guidance only. For batch specific data refer to the Certificate of Analysis.
Solubility
| Solvent | Max Conc. mg/mL | Max Conc. mM | |
|---|---|---|---|
| Solubility | |||
| DMF | 29.69 | 50 | |
| DMSO | 2.97 | 5 |
Preparing Stock Solutions
for Janelia Fluor® 646, NHS ester
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 all 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 |
Calculators
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Background References
References are publications that support the biological activity of the product.
- Zheng Rational design of fluorogenic and spontaneously blinking labels for super-resolution imaging. ACS Cent.Sci. 2019 PMID: 31572787
- Ticau Single-molecule studies of origin licensing reveal mechanisms ensuring bidirectional helicase loading. Cell 2015 PMID: 25892223
- Hong 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. 2009 PMID: 19666497
- Deng CASFISH: CRISPR/Cas9-mediated in situ labeling of genomic loci in fixed cells. Proc.Natl.Acad.Sci.USA. 2015 PMID: 26324940
- Grimm A general method to improve fluorophores for live-cell and single-molecule microscopy. Nat.Methods 2015 PMID: 25599551
- Legant High-density three-dimensional localization microscopy across large volumes. Nat.Methods 2016 PMID: 26950745
- Li Real-time imaging of Huntingtin aggregates diverting target search and gene transcription. Elife 2016 PMID: 27484239
- Schmidt Live cell imaging reveals the dynamics of telomerase recruitment to telomeres. Cell 2016 PMID: 27523609
Product Specific Notices for Janelia Fluor® 646, NHS ester
For research use only
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