Janelia Fluor® 525 NHS Ester (Succinimidyl Ester)
(HB8455)
Product overview
| Name | Janelia Fluor® 525 NHS Ester (Succinimidyl Ester) |
| Biological description | Cell-permeable, yellow fluorescent dye with an NHS ester (succinimidyl ester (SE)) reactive group. NHS esters react with primary amines on proteins and are commonly used for conjugating dyes to proteins, antibodies amine-modified oligonucleotides etc. Suitable for super resolution microscopy (SRM) including techniques such as dSTORM (both live and fixed cells), cellular imaging when combined with the HaloTag or SNAP-tag self labelling systems and confocal microscopy. 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 primary amine 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 |
| Storage of solutions | Prepare and use solutions on the same day if possible. Store solutions at -20°C for up to one month if storage is required. Equilibrate to RT and ensure the solution is precipitate free before use. |
| Shipping Conditions | Stable for ambient temperature shipping. Follow storage instructions on receipt. |
| 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 NHS Ester (Succinimidyl Ester)
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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