Ready-to-use for application in fluorescence microscopy procedures. With increasing age, the autofluorescent pigment lipofuscin accumulates in the cytoplasm of many cell types, including neurons. The presence of lipofuscin granules can complicate the use of fluorescence microscopy in the central nervous system because of its broad excitation and emission spectra, which overlaps with those of most commonly used fluorophores. The Autofluorescence Eliminator Reagent will reduce or eliminate lipofuscin-like autofluorescence without adversely affecting other fluorescent label in sections of human, monkey or rat neural tissue as well as other tissues.
Fluorescence immunohistochemistry counterstain.
1.After immunofluorescence histochemistry, the sections are immersed in PBS for 5 min.·
2.Sections are immersed in 70% ethanol for 5 min.·
3.Sections are immersed in Autofluorescence Eliminator Reagent for 5 min.
4.Sections are immersed in three changes of 70% ethanol for 1 min. each time.·
5.Sections are mounted using an antifading solution (e.g. DABCO, non-xylene-based).
Note: Reagent will block all fluorescence if incubation is too long; length of incubation time and number of 70% ethanol washes can be altered and optimized for various tissues and fluorescent antibody probes. In some instances the reagent can be applied prior to immunofluorescence, however in most cases better results are obtained when used as we direct above (use after immunofluorescence procedures are completed).
Optimal time of incubation with the Autofluorescence Eliminator Reagent should be determined by the end user for each sample.
Absorption maximum: 595-605
Liquid in 70% ethanol containing no preservatives. Store at room temperature in the dark.
Storage and Stability
When stored at room temperature, this reagent is stable up to the expiration date printed on the label. Do not freeze or expose to elevated temperatures.
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