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Submit ReviewPA Janelia Fluor® 646, NHS ester
Description: Red photoactivatable dye (Reactive group: NHS ester). Application: super resolution microscopy (SRM), single molecule tracking and live cell imaging
Chemical Name: 2,5-Dioxo-1-pyrrolidinyl 3,7-di-1-azetidinyl-2'-diazo-5,5-dimethyl-2',3'-dihydro-5H-3'-oxospiro[dibenzo[b,e]siline-10,1'-indene]-6'-carboxylate
Purity: ≥90% (HPLC)
Biological Activity for PA Janelia Fluor® 646, NHS ester
Key information: PA Janelia Fluor® 646, NHS ester is a photoactivatable fluorescent dye; supplied with an NHS ester reactive group for the labeling of primary amines. Suitable for live and fixed cell imaging.
Application: Suitable for single molecule tracking and super resolution microscopy in live cells, specifically sptPALM (live cells) and PALM (fixed cells) techniques.
Properties and Photophysical Data: Non-fluorescent until activated at 405 nm. NHS ester can be converted to relevant substrate for use in self-labeling tag systems, e.g. HaloTag® and SNAP-tag®. Can be multiplexed with PA Janelia Fluor® 549, NHS ester (Cat. No. 6149) to perform two-color sptPALM in live cells with activation at 405nm. Upon protein conjugation the efficiencies of photoconversion improve substantially. Low background staining. Excitation and emission maxima (λ) are 649 - 651 nm and 663 - 665 nm, respectively.
Please see the protocol for further information on protein/antibody labeling and conjugation.
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 PA Janelia Fluor® 646, SE.
Licensing Information
Sold under license from the Howard Hughes Medical Institute, Janelia Research Campus
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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 PA Janelia Fluor® 646, NHS ester
| M. Wt | 617.74 |
| Formula | C34H31N5O5Si |
| Storage | Store at -20°C |
| Purity | ≥90% (HPLC) |
| CAS Number | 2093120-32-0 |
| PubChem ID | 127021042 |
| InChI Key | XVIATAHKOZWKHH-UHFFFAOYSA-N |
| Smiles | O=C(ON1C(CCC1=O)=O)C2=CC=C(C(C3=[N+]=[N-])=O)C(C43C5=CC=C(N6CCC6)C=C5[Si](C)(C)C7=CC(N8CCC8)=CC=C47)=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 PA Janelia Fluor® 646, NHS ester
| Solvent | Max Conc. mg/mL | Max Conc. mM | |
|---|---|---|---|
| Solubility | |||
| DMSO | 12.35 | 20 |
Preparing Stock Solutions for PA Janelia Fluor® 646, NHS ester
The following data is based on the product molecular weight 617.74. 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 |
|---|---|---|---|
| 0.2 mM | 8.09 mL | 40.47 mL | 80.94 mL |
| 1 mM | 1.62 mL | 8.09 mL | 16.19 mL |
| 2 mM | 0.81 mL | 4.05 mL | 8.09 mL |
| 10 mM | 0.16 mL | 0.81 mL | 1.62 mL |
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Product Datasheets for PA Janelia Fluor® 646, NHS ester
References for PA Janelia Fluor® 646, NHS ester
References are publications that support the biological activity of the product.
Grimm et al (2016) Bright photoactivatable fluorophores for single-molecule imaging. Nat.Methods 13 985 PMID: 27776112
Li et al (2016) Real-time imaging of Huntingtin aggregates diverting target search and gene transcription. eLife 5 e17056 PMID: 27484239
If you know of a relevant reference for PA Janelia Fluor® 646, NHS ester, please let us know.
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3 Citations for PA Janelia Fluor® 646, NHS ester
Citations are publications that use Tocris products. Selected citations for PA Janelia Fluor® 646, NHS ester include:
Jeroen et al (2022) Microscopy-based single-cell proteomic profiling reveals heterogeneity in DNA damage response dynamics. Cell Rep Methods 2 100237 PMID: 35784653
Genshaft et al (2021) Live cell tagging tracking and isolation for spatial transcriptomics using photoactivatable cell dyes Nat Commun 12 4995 PMID: 34404785
Sheila S et al (2019) Visualizing Transcription Factor Binding on Mitotic Chromosomes Using Single-Molecule Live-Cell Imaging. Methods Mol Biol 2038 239-250 PMID: 31407289
Do you know of a great paper that uses PA Janelia Fluor® 646, NHS ester from Tocris? Please let us know.
Reviews for PA Janelia Fluor® 646, NHS ester
Average Rating: 5 (Based on 2 Reviews.)
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BG labelled PA JF646.
By
Abigail Walker on 05/21/2024
Assay Type: In Vitro
Species: Human
Cell Line/Tissue: HEK 293 cells
We conjugated PA JF656 with the SNAP label BG, and used it for single molecule imaging in HEK 293 cells (both live and fixed). The dye works very well, it is stable, bright, and activates and bleaches well (better than other photoactivatable dyes I have used).We have used it for imaging at 1-10 nM as we found some non-specific binding at concentrations higher than this, but potentially this could be removed with additional wash steps.
PA-JF646 for tracking single recombinant proteins.
By
Anonymous on 02/26/2018
Assay Type: In Vitro
Species: Bacteria
This product was used to label recombinant proteins and image them with single molecule sensitivity. The degree of labeling achieved was 1.4 (average). The photoactivation efficiency of the dye is excellent and can be used to track a few molecules at a time, for long periods. The dye is highly photostable and can be used to track proteins for ~10-15 seconds under ideal tracking conditions. I attach here an image before and after photoactivation of the dye. Upon photoactivation, single dye molecules conjugated to the protein are visible.
Protocols for PA Janelia Fluor® 646, NHS ester
The following protocol features additional information for the use of PA Janelia Fluor® 646, NHS ester (Cat. No. 6150).
Literature in this Area
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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