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AG310 | Fluoro-Jade® B (degenerating neurons)

50 mg  
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      Replacement Information

      Key Specifications Table

      Key Applications
      Catalogue NumberAG310
      Brand Family Chemicon®
      Trade Name
      • Fluoro-Jade
      • Chemicon
      DescriptionFluoro-Jade® B (degenerating neurons)
      OverviewFluoro-Jade® B is a polyanionic fluorescein derivitave which sensitively and specifically binds to degenerating neurons. It is a dark red powder that has a green iridescence with excitation peak at 480 nm and emission peak at 525 nm. The filter used for visualizing Fluoro-Jade® B is a fluorescein/FITC filter. Fluoro-Jade® B can be used on most tissue section types and thicknesses including frozen, vibratomed cryostat or paraffin-embedded sections from 3-50 μm. Fluoro-Jade® B is faster and more reliable than older methods (e.g. suppressed silver) for the unequivocal qualitative detection and quantitative measurement of both gross and fine scale neuronal degeneration. Fluoro-Jade B will not distinguish between apoptotic and necrotic neurons. Astrocytic labeling is occasionally observed with Fluoro-Jade B, especially in chronic studies. It is believed that such labelling represents astrocytic activation and not death as is the case for neurons (see Ye X et. al. Brain Res. Prot. 8 (2001) 104-112; Anderson, KJ (2003) J Neurotrauma 20(11):1223-31.). The specific protein or lipid component labeled by Fluoro-Jade B is not known.

      APPEARANCE: Dark red powder with green iridescence.


      EXCITATION PEAK: 480 nm

      EMISSION PEAK: 525 nm

      FILTER SYSTEM: Fluorescein / FITC

      SOLUBILITY: Very soluble in water and bases, moderately soluble in alcohol and weak acids.

      TOXICITY: Although the compound appears to be of low toxicity, it has not been extensively evaluated and therefore routine laboratory caution should be exercised. Not intended for human consumption.
      Product Information
      Key Applications
      • Immunohistochemistry


      Following appropriate survival interval, animals were perfused with 300 mL of 0.1 M neutral phosphate buffered 10% formalin (4% formaldehyde) via the ascending aorta, while clamping off the descending aorta. The brains were postfixed at least overnight in the same fixative solution plus 20% sucrose. Tissue was cut on a freezing sliding microtome at a thickness of 25 um. The sections were collected in 0.1 M neutral phosphate buffer. The sections were typically mounted on 2% gelatin coated slides and then air dried on a slide warmer at 50 degrees C for at least half an hour. The slides were first immersed in a solution containing 1% sodium hydroxide in 80% alcohol (20 mL of 5% NaOH added to 80 mL absolute alcohol) for 5 minutes. This was followed by 2 minutes in 70% alcohol and 2 minutes in distilled water. The slides were then transferred to a solution of 0.06% potassium permanganate for 10 minutes, preferably on a shaker table to insure consistent background suppression between sections. The slides were then rinsed in distilled water for 2 minutes. The staining solution was prepared from a 0.01% stock solution for Fluoro-Jade® B that was made by adding 10 mg of the dye powder to 100 mL of distilled water. To make up 100 mL of staining solution, 4 mL of the stock solution was added to 96 mL of 0.1% acetic acid vehicle. This results in a final dye concentration of 0.0004%. The stock solution, when stored in the refrigerator was stable for months, whereas the staining solution was typically prepared within 10 minutes of use and was not reused. After 20 minutes in the staining solution, the slides were rinsed for one minute in each of three distilled water washes. Excess water was removed by briefly (about 15 s) draining the slides vertically on a paper towel. The slides were then placed on a slide warmer, set at approximately 50 degrees C, until they were fully dry, (eg. 5-10 min). The dry slides were cleared by immersion in xylene for at least a minute before coverslipping with DPX (Fluka, Milwaukee WI, or Sigma Chem. Co., St. Louis, MO), a non-aqueous non-fluorescent plastic mounting media.


      The tissue was then examined using an epifluorescent microscope with blue (450-490 nm) excitation light. A barrier filter that allows passage of all wavelengths longer than 515 nm will result in a yellow-green emission color, where as a notch filter, (eg. 515-565 nm) will result in a green emission color. Most filters designed for visualizing fluorescein or FITC (eg. the Nikon B-2A or the B-3A filter cubes) will be suitable for visualizing Fluoro-Jade® B.

      Frequently Asked Questions:

      1) What can be done if the background level is too high relative to specific staining?

      Answer: Leave in fresh potassium permanganate longer, about twenty minutes, or dilute the Fluoro-Jade® concentration by half, Fluoro-Jade® B (.0002%)

      2) Can Fluoro-Jade® be used with paraffin processed tissue?

      Answer: Yes. Use xylene to remove paraffin, then rinse twice with alcohol. Also works with cryostat cut unfixed tissue.

      3) Can it be combined with immunofluorescence?

      Answer Yes. Although, sometimes pretreatment procedures can attenuate immunofluorescent labeling. If so, the time in potassium permanganate solution should be reduced as necessary. Dye concentration may also need to be reduced.
      Biological Information
      PurityThin layer chromatography using cellulose plates and a solvent system of n-propinol, water, and ammonium hydroxide (6:5:2) revealed the presence of two fluorescent isomers and two trace non-fluorescent bands. No amount of fluorescein or Fluoro-Jade® was present.
      Physicochemical Information
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Usage Statement
      • Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
      Storage and Shipping Information
      Storage ConditionsThe lyophilized powder should be stored well sealed at room temperature, preferable in a desiccator (due to its hygroscopic nature), for up to one year from date of receipt. The liquid stock solution (0.01%) in distilled water can be stored at 2-8°C for up to 3 months. The working solution (0.0004%) in 0.1% acetic acid should be prepared fresh and not be stored or reused.
      Packaging Information
      Material Size50 mg
      Transport Information
      Supplemental Information


      Certificates of Analysis

      TitleLot Number
      FLUORO-JADE#174; B - 2115915 2115915
      FLUORO-JADE#174; B - 2020489 2020489
      FLUORO-JADE#174; B - 2043869 2043869


      Reference overviewApplicationSpeciesPub Med ID
      Neuregulin-1 is neuroprotective in a rat model of organophosphate-induced delayed neuronal injury.
      Li, Y; Lein, PJ; Liu, C; Bruun, DA; Giulivi, C; Ford, GD; Tewolde, T; Ross-Inta, C; Ford, BD
      Toxicology and applied pharmacology 262 194-204 2012

      Show Abstract
      22583949 22583949
      An experimental protocol for mimicking pathomechanisms of traumatic brain injury in mice.
      Christiane Albert-Weißenberger,Csanád Várrallyay,Furat Raslan,Christoph Kleinschnitz,Anna-Leena Sirén,Christiane Albert-Wei,Csan V,Anna-Leena Sir
      Experimental & translational stroke medicine 4 2012

      Show Abstract
      22300472 22300472
      Vulnerability of postnatal hippocampal neurons to seizures varies regionally with their maturational stage.
      Maria-Leonor Lopez-Meraz,Claude G Wasterlain,Luisa L Rocha,Suni Allen,Jerome Niquet
      Neurobiology of disease 37 2010

      Show Abstract Full Text Article
      19879360 19879360
      Allele-specific RNA silencing of mutant ataxin-3 mediates neuroprotection in a rat model of Machado-Joseph disease.
      Alves, Sandro, et al.
      PLoS ONE, 3: e3341 (2008) 2008

      Show Abstract
      Immunoblotting (Western)Rat18841197 18841197
      Differential neuroprotective properties of endogenous and exogenous erythropoietin in a mouse model of traumatic brain injury.
      Na'ama A Shein,Nikolaos Grigoriadis,Alexander G Alexandrovich,Constantina Simeonidou,Evangelia Spandou,Jeanna Tsenter,Ido Yatsiv,Michal Horowitz,Esther Shohami
      Journal of neurotrauma 25 2008

      Show Abstract
      18260794 18260794
      Status epilepticus induces time-dependent neuronal and astrocytic expression of interleukin-1 receptor type I in the rat limbic system.
      Ravizza, T and Vezzani, A
      Neuroscience, 137: 301-8 (2006) 2006

      Show Abstract
      16289587 16289587
      Neuropathology and neurodegeneration in rodent brain induced by lentiviral vector-mediated overexpression of alpha-synuclein.
      Lauwers, Erwin, et al.
      Brain Pathol., 13: 364-72 (2003) 2003

      Show Abstract
      12946025 12946025
      Fluoro-Jade B: a high affinity fluorescent marker for the localization of neuronal degeneration.
      Schmued, L C and Hopkins, K J
      Brain Res., 874: 123-30 (2000) 2000

      Show Abstract
      10960596 10960596

      Data Sheet


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