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Submit ReviewJanelia Fluor® 635, NHS ester
Description: Red dye (Reactive group: NHS ester). Application: flow cytometry, confocal microscopy, super resolution microscopy. Suitable for live cell imaging
Alternative Names: JF 635,NHS
Chemical Name: 1-[10-[2-Carboxy-5-[[(2,5-dioxo-1-pyrrolidinyl)oxy]carbonyl]phenyl]-9,9-dimethyl-7-[1-(3-fluoroazetidinyl)]-9-silaanthracen-2(9H)-ylidene]-3-fluoroazetidinium, inner salt
Purity: ≥95% (HPLC)
Biological Activity for Janelia Fluor® 635, NHS ester
Key Information: Janelia Fluor® 635, 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). 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 635 nm and 652 nm, respectively; quantum yield = 0.56; extinction coefficient = 167,000 M-1cm-1 (measured in ethanol or TFE plus 0.1% TFA); A280 correction factor = 0.0524; lactone-zwitterion equilibrium constant (KL-Z) <0.0001. Exhibits low nonspecific background staining.
Please see the protocol for further information on protein/antibody labeling and conjugation.
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.
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® 635, SE.
Scientific Data
Confocal Microscopy
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Applications on mitochondria for Janelia Fluor® 635, NHS ester. Application of Janelia Fluor® 635, NHS ester dye on mitochondria; Left: Mitochondria were labeled with anti-TOM20 primary antibodies. The secondary antibodies were conjugated to JF 635 dye. Maximum intensity projection of confocal z-stack, recorded on an Olympus FV1200 confocal microscope. Right: Enlarged portion of left-hand image. Images kindly provided by Dr Kirstin Elgass, Monash University.
Licensing Information
Sold under license from the Howard Hughes Medical Institute, Janelia Research Campus
Optical Data for Janelia Fluor® 635, NHS ester
| Emission Color | Red |
|---|---|
| Brightness | |
| λabs | 635 nm |
| λem | 652 nm |
| Extinction Coefficient (ε) | 167,000 M-1cm-1 |
| Quantum Yield (φ) | 0.56 |
| Closest Laser line | 635 nm |
| Reactive Group | NHS ester |
| Reactivity | Primary amines |
| Cell Permeable | Yes |
| Correction Factor 280 | 0.0524 |
| Application | Confocal Microscopy, Super Resolution Microscopy, Flow Cytometry |
| Alternative For | Alexa Fluor® 633, Atto 633, CF633 |
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.
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Technical Data for Janelia Fluor® 635, NHS ester
| M. Wt | 629.69 |
| Formula | C33H29F2N3O6Si |
| Storage | Store at -20°C |
| Purity | ≥95% (HPLC) |
| PubChem ID | 132470914 |
| InChI Key | YVKNNZJUJGADIJ-UHFFFAOYSA-N |
| Smiles | O=C(ON1C(CCC1=O)=O)C2=CC=C(C([O-])=O)C(C(C3=CC=C(N4CC(F)C4)C=C3[Si]5(C)C)=C(C=C/6)C5=CC6=[N+]7CC(F)C\7)=C2 |
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.
Solubility Data for Janelia Fluor® 635, NHS ester
| Solvent | Max Conc. mg/mL | Max Conc. mM | |
|---|---|---|---|
| Solubility | |||
| DMSO | 62.97 | 100 |
Preparing Stock Solutions for Janelia Fluor® 635, NHS ester
The following data is based on the product molecular weight 629.69. 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 |
|---|---|---|---|
| 1 mM | 1.59 mL | 7.94 mL | 15.88 mL |
| 5 mM | 0.32 mL | 1.59 mL | 3.18 mL |
| 10 mM | 0.16 mL | 0.79 mL | 1.59 mL |
| 50 mM | 0.03 mL | 0.16 mL | 0.32 mL |
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Product Datasheets for Janelia Fluor® 635, NHS ester
References for Janelia Fluor® 635, NHS ester
References are publications that support the biological activity of the product.
Grimm et al (2017) A general method to fine-tune fluorophores for live-cell and in vivo imaging. Nat.Methods 14 987 PMID: 28869757
Zheng et al (2019) Rational design of fluorogenic and spontaneously blinking labels for super-resolution imaging. ACS Cent.Sci. 5 1602 PMID: 31572787
If you know of a relevant reference for Janelia Fluor® 635, NHS ester, please let us know.
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Protocols for Janelia Fluor® 635, NHS ester
The following protocol features additional information for the use of Janelia Fluor® 635, NHS ester (Cat. No. 6419).
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Fluorescent Dyes and Probes Research Product GuideUpdated
This product guide provides a background to the use of Fluorescent Dyes and Probes, as well as a comprehensive list of our:
- Fluorescent Dyes, including Janelia Fluor® Dyes
- Fluorescent Probes for Advanced Microscopy and Live Cell Imaging
- Fluorescent Probes to Support In Vivo, Deep Tissue, and Bioluminescence Imaging
- Fluorescent Probes and Reagents for Organoids and 3D Cell Culture Imaging
- Aptamer-based RNA Imaging Reagents
- Fluorescent Probes for Imaging Bacteria
- TSA VividTM Fluorophore Kits
- TSA Reagents for Enhancing IHC, ICC & FISH Signals
- Tissue Clearing Kits and Reagents