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Submit ReviewPA Janelia Fluor® 646, SE
Description: Red photoactivatable dye (Reactive group: NHS ester). Applications: super resolution microscopy (PALM), 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, SE
PA Janelia Fluor® 646, SE is a photoactivatable fluorescent dye; supplied as an NHS ester for coupling to primary amine groups. Non-fluorescent until activated at 365 nm. NHS ester can be converted to relevant substrate for use in self-labeling tag systems, e.g. HaloTag® and SNAP-tag®. Suitable for single molecule tracking and super resolution microscopy in live cells, specifically live cell sptPALM and fixed cell PALM. Can be multiplexed with PA Janelia Fluor® 549, SE (Cat. No. 6149) to perform two-colour sptPALM in live cells with activation at 405nm. Cell permeable. Excitation maximum = 649 - 651 nm; emission maximum = 663 - 665 nm. Upon protein conjugation the efficiencies of photoconversion improve substantially. Low background staining.
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.
Licensing Information
Sold under license from the Howard Hughes Medical Institute, Janelia Research Campus
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Technical Data for PA Janelia Fluor® 646, SE
| 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, SE
| Solvent | Max Conc. mg/mL | Max Conc. mM | |
|---|---|---|---|
| Solubility | |||
| DMSO | 12.35 | 20 |
Preparing Stock Solutions for PA Janelia Fluor® 646, SE
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, SE
References for PA Janelia Fluor® 646, SE
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, SE, please let us know.
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3 Citations for PA Janelia Fluor® 646, SE
Citations are publications that use Tocris products. Selected citations for PA Janelia Fluor® 646, SE include:
Genshaft et al (2021) Live cell tagging tracking and isolation for spatial transcriptomics using photoactivatable cell dyes Nat Commun 12 4995 PMID: 34404785
Jeroen et al (2022) Microscopy-based single-cell proteomic profiling reveals heterogeneity in DNA damage response dynamics. Cell Rep Methods 2 100237 PMID: 35784653
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
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Reviews for PA Janelia Fluor® 646, SE
Average Rating: 5 (Based on 1 Review.)
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PA-JF646 for tracking single recombinant proteins.
By
Anonymous on 02/26/2018
Assay Type: In Vitro
Species: Other
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, SE
The following protocol features additional information for the use of PA Janelia Fluor® 646, SE (Cat. No. 6150).
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Fluorescent Dyes and Probes Research Product Guide
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