Janelia Fluor® 549, SE
(HB7336)
Product overview
| Name | Janelia Fluor® 549, SE |
| Biological description | Cell permeable, yellow fluorescent dye supplied as an NHS ester / succinimidyl ester (SE) for coupling to primary amine (NHS) groups. NHS esters are used to label the primary amines of proteins and are commonly used for conjugating dyes to a protein or antibody. May be used for cellular imaging when combined with the HaloTag or SNAP-tag self labelling systems. Suitable for confocal microscopy and super resolution microscopy (SRM) including techniques such as dSTORM (both live and fixed cells) and STED. Also suitable for flow cytometry. Janelia Fluor® 549 is 2 x brighter than TMR and Cy3 in vitro and live-cell experiments.
|
| Alternative names | JF549, JF549-SE |
| Description | Yellow dye for coupling to NHS groups. Suitable for dSTORM, STED, confocal microscopy, live cell imaging and flow cytometry. |
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Images
Fig 1. ß-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 1. β-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody. HeLa cells were stained using FITC Phalloidin (HB0814) and a mouse monoclonal β-tubulin (HB6491) antibody from Hello Bio which was visualised using a Janelia Fluor® 549 SE conjugated polyclonal goat anti-mouse antibody. For more details see protocols #1 and #2 in the application notes below.
Fig 2. GFAP expression in rat brain visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 2. GFAP expression in rat brain visualised using a Janelia Fluor® 549 conjugated secondary antibody. Rat horizontal brain sections were stained using mouse monoclonal GFAP (HB8267) and Neurofilament L (HB7266) antibodies from Hello Bio which were visualised using a Janelia Fluor® 549 SE conjugated goat anti-mouse antibody and a DyLight 488 conjugated goat anti- rabbit antibody. For more details see protocols #1 and #3 in the application notes below.
Fig 3. ß-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 3. β-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody. HeLa cells were stained using FITC Phalloidin (HB0814) and a mouse monoclonal β-tubulin antibody (HB6491) from Hello Bio which was visualised using a Janelia Fluor® 549 SE conjugated polyclonal goat anti- mouse antibody. For more details see protocols #1 and #2 in the application notes
Figure 4. Janelia Fluor® 549 conjugated secondary antibodies show superior antifade performance compared to those conjugated with DyLight 550
Fig 1. ß-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 1. β-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody. HeLa cells were stained using FITC Phalloidin (HB0814) and a mouse monoclonal β-tubulin (HB6491) antibody from Hello Bio which was visualised using a Janelia Fluor® 549 SE conjugated polyclonal goat anti-mouse antibody. For more details see protocols #1 and #2 in the application notes below.
Fig 2. GFAP expression in rat brain visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 2. GFAP expression in rat brain visualised using a Janelia Fluor® 549 conjugated secondary antibody. Rat horizontal brain sections were stained using mouse monoclonal GFAP (HB8267) and Neurofilament L (HB7266) antibodies from Hello Bio which were visualised using a Janelia Fluor® 549 SE conjugated goat anti-mouse antibody and a DyLight 488 conjugated goat anti- rabbit antibody. For more details see protocols #1 and #3 in the application notes below.
Fig 3. ß-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody.
Figure 3. β-tubulin expression in HeLa cells visualised using a Janelia Fluor® 549 conjugated secondary antibody. HeLa cells were stained using FITC Phalloidin (HB0814) and a mouse monoclonal β-tubulin antibody (HB6491) from Hello Bio which was visualised using a Janelia Fluor® 549 SE conjugated polyclonal goat anti- mouse antibody. For more details see protocols #1 and #2 in the application notes
Figure 4. Janelia Fluor® 549 conjugated secondary antibodies show superior antifade performance compared to those conjugated with DyLight 550
Optical Data
| Fluorescence spectra |  |
| Reactivity | Primary amines |
| Emission color | Yellow |
| Max excitation wavelength | 549 nm |
| Max emission wavelength | 571 nm |
| Closest laser lines | 532nm |
| Spectrally similar dyes | Alexa Fluor® 546 |
| Quantum Yield (φ) | 0.88 |
| Extinction Coefficient (ε) | 101,000 M-1cm-1 |
| Cell permeable | Yes |
| Reactive Group | NHS ester |
Biological Data
| Application notes | #Protocol 1: Conjugation of Janelia© Fluor 549 SE to antibodies
#Protocol 2: Immunocytochemistry
#Protocol 3: Immunohistochemistry
|
Solubility & Handling
| Storage instructions | -20°C |
| Solubility overview | Soluble in DMSO |
| Handling | This compound is light and temperature sensitive; exposure to light may affect compound performance. We therefore recommend storing the material in a freezer and protecting from light. |
| Shipping conditions | Ship on ice |
| Important | This product is for RESEARCH USE ONLY and is not intended for therapeutic or diagnostic use. Not for human or veterinary use |
Chemical Data
| Chemical name | 3,6-Di-1-azetidinyl-9-[2-carboxy-5-[[(2,5-dioxo-1-pyrrolidinyl)oxy]carbonyl]phenyl]xanthylium, inner salt |
| Molecular Weight | 551.5 |
| Chemical structure | ![Janelia Fluor® 549, SE [1811539-32-8] Chemical Structure](https://hellobio.com/media/catalog/product//h/b/hb7336.png "Janelia Fluor® 549, SE [1811539-32-8]") |
| Molecular Formula | C31H25N3O7 |
| CAS Number | 1811539-32-8 |
| PubChem identifier | 124201856 |
| SMILES | C1CN(C1)C2=CC3=C(C=C2)C(=C4C=CC(=[N+]5CCC5)C=C4O3)C6=C(C=CC(=C6)C(=O)ON7C(=O)CCC7=O)C(=O)[O-] |
| Source | Synthetic |
| InChiKey | DWUIXAGWHCMXHN-UHFFFAOYSA-N |
| Licensing details | Sold under license from the Howard Hughes Medical Institute, Janelia Research Campus |
References for Janelia Fluor® 549, SE
References are publications that support the biological activity of the product
-
Rational Design of Bioavailable Photosensitizers for Manipulation and Imaging of Biological Systems.
Binns TC et al (2020) Cell chemical biology 27 : 1063-1072.e7 -
A general method to fine-tune fluorophores for live-cell and in vivo imaging.
Grimm JB et al (2017) Nature methods 14 : 987-994 -
Synthesis of Janelia Fluor HaloTag and SNAP-Tag Ligands and Their Use in Cellular Imaging Experiments.
Grimm JB et al (2017) Methods in molecular biology (Clifton, N.J.) 1663 : 179-188 -
A general method to improve fluorophores for live-cell and single-molecule microscopy.
Grimm JB et al (2015) Nature methods 12 : 244-50, 3 p following 250
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