Janelia Fluor® 525, free acid

Technical documents: CoA Datasheet

Product overview

Name Janelia Fluor® 525, free acid
Biological description

Cell-permeable, yellow fluorescent dye supplied as a free acid. 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


Yellow dye supplied as a free acid. Suitable for super resolution microscopy (e.g. dSTORM), confocal microscopy and live cell imaging.

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Fig1. Excitation and emission spectra of Janelia Fluor ® 525, free acid.

Figure1. Excitation and emission spectra of Janelia Fluor® 525, free acid. 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 1 in application notes below.

Optical Data

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 Free acid

Biological Data

Application notes

#Protocol 1: Measurement of excitation and emission spectra of Janelia Fluor ® 525, free acid

  • Janelia Fluor ® 525, free acid was prepared at 1µm in PBS.
  • Spectra were generated on a Tecan Infinite M200 PRO using the following parameters:
    • Excitation: Recording at 618nm while exciting between 280nm and 590nm
    • Emission: Exciting at 484nm while recording between 510nm and 800nm
    • Absorbance: Measured between 300 and 800nm

Solubility & Handling

Storage instructions -20°C
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



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Chemical Data

Chemical name 3,6-Di-1-(3,3-difluoroazetidinyl)-9-[2,5-dicarboxy-phenyl]xanthylium, inner salt
Molecular Weight 526.44
Chemical structure  Janelia Fluor® 525, free acid Chemical Structure
Molecular Formula C27H19F4N2O5
SMILES O=C(O)c6cc(C=1c4ccc(cc4OC2=CC(\C=CC=12)=[N+]3/CC(F)(F)C3)N5CC(F)(F)C5)c(cc6)C([O-])=O
Source Synthetic
Licensing details

Sold under license from the Howard Hughes Medical Institute, Janelia Research Campus

References for Janelia Fluor® 525, free acid

References are publications that support the biological activity of the product
  • 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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