Janelia Fluor® 635, NHS ester
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%
Biological Activity
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 with our sister company Novus Biologicals.
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
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Applications on mitochondria for Janelia Fluor® 635, SE. Application of Janelia Fluor® 635, SE 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.
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.
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
-
A general method to fine-tune fluorophores for live-cell and in vivo imaging.
Grimm et al.
Nat.Methods, 2017;14:987 -
Rational design of fluorogenic and spontaneously blinking labels for super-resolution imaging.
Zheng et al.
ACS Cent.Sci., 2019;5:1602
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