Janelia Fluor® 525, SE
(HB8455)
Product overview
| Name | Janelia Fluor® 525, 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). Can also be multiplexed with Janelia Fluor ® 635 SE for two color imaging. Spectrally similar dyes: Alexa Fluor® 532, Alexa Fluor® 514, Atto 532, CF514, CF532 |
| Alternative names | JF525,SE |
| Description | Yellow dye for coupling to NHS groups. Suitable for super resolution microscopy (e.g. dSTORM), confocal microscopy and live cell imaging. |
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Images
Fig 1. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody.
Figure 1. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody. Rat horizontal brain sections were stained using mouse monoclonal GFAP (HB8267) and rabbit monoclonal 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 #2 in the application notes below.
Fig 2. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody.
Figure 2. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody. Rat horizontal brain sections were stained using mouse monoclonal GFAP (HB8267) and rabbit monoclonal 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 #2 in the application notes below.
Fig 3. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody.
Figure 3. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody. Rat horizontal brain sections were stained using mouse monoclonal GFAP (HB8267) and rabbit monoclonal 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 #2 in the application notes below.
Fig4. Excitation and emission spectra of Janelia Fluor ® 525, SE.
Figure4. Excitation and emission spectra of Janelia Fluor ® 525, SE. Janelia Fluor® 525 is a yellow fluorescent dye suitable for confocal and super resolution microscopy. A 1 µM solution was prepared in PBS before excitation, emission and absorbance spectra were generated. For protocol see #Protocol 3 in application notes below.
Fig 1. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody.
Figure 1. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody. Rat horizontal brain sections were stained using mouse monoclonal GFAP (HB8267) and rabbit monoclonal 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 #2 in the application notes below.
Fig 2. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody.
Figure 2. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody. Rat horizontal brain sections were stained using mouse monoclonal GFAP (HB8267) and rabbit monoclonal 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 #2 in the application notes below.
Fig 3. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody.
Figure 3. GFAP expression in rat brain visualised using a Janelia Fluor® 525 conjugated secondary antibody. Rat horizontal brain sections were stained using mouse monoclonal GFAP (HB8267) and rabbit monoclonal 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 #2 in the application notes below.
Fig4. Excitation and emission spectra of Janelia Fluor ® 525, SE.
Figure4. Excitation and emission spectra of Janelia Fluor ® 525, SE. Janelia Fluor® 525 is a yellow fluorescent dye suitable for confocal and super resolution microscopy. A 1 µM solution was prepared in PBS before excitation, emission and absorbance spectra were generated. For protocol see #Protocol 3 in application notes below.
Optical Data
| Reactivity | Primary amines |
| Emission color | Yellow |
| Max excitation wavelength | 525 nm |
| Max emission wavelength | 549 nm |
| Quantum Yield (φ) | 0.91 |
| Extinction Coefficient (ε) | 122,000 M-1cm-1 |
| Cell permeable | Yes |
| Reactive Group | NHS ester |
Biological Data
| Application notes | #Protocol 1: Conjugation of Janelia© Fluor 525 SE to antibodies
#Protocol 2: Immunohistochemistry
#Protocol 3: Measurement of excitation and emission spectra of Janelia Fluor ® 525, SE
|
Solubility & Handling
| Storage instructions | -20° |
| Solubility overview | Soluble in DMSO |
| 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-(3,3-difluoroazetidinyl)-9-[2-carboxy-5-[[(2,5-dioxo-1-pyrrolidinyl)oxy]carbonyl]phenyl]xanthylium, inner salt |
| Molecular Weight | 623.51 |
| Chemical structure |  |
| Molecular Formula | C31H21F4N3O7 |
| PubChem identifier | 134160239 |
| SMILES | O=C(ON1C(CCC1=O)=O)C2=CC=C(C([O-])=O)C(C(C3=CC=C(N4CC(F)(F)C4)C=C3O5)=C(C=C/6)C5=CC6=[N+]7CC(F)(F)C\7)=C2 |
| Source | Synthetic |
| InChiKey | VPHPPEABRFKVDC-UHFFFAOYSA-N |
| Licensing details | Sold under license from the Howard Hughes Medical Institute, Janelia Research Campus |
References for Janelia Fluor® 525, SE
References are publications that support the biological activity of the product
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A general method to fine-tune fluorophores for live-cell and in vivo imaging.
Grimm JB et al (2017) Nature methods 14 : 987-994
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