6-Aminofluorescein

6-Aminofluorescein (6-AF) is a fluorescent labeling reagent. 6-Aminofluorescein can be used to label fullerene-based liposome nanostructures termed ‘buckysomes′ (Ex/Em = 490/515 nm)^[1]. In Vitro:Guide (The following is our recommended protocol. This protocol is only a guide and should be modified according to your specific needs).
Buckysomes are coupled to 6-Aminofluorescein
400 μL of buckysomes (2 mg/mL) is incubated with 100 μL each of 0.25 M EDC (1-Ethyl-3-(3-dimethylaminopropyl)carbodiimide (HY-D0178) and 0.25 M sulfo-NHS for 2 hrs at room temperature.
The pH is adjusted to 7.0 using NaOH.
300 μL of 6-Aminofluorescein (1 mg/mL prepared in DMSO) is added and incubated overnight at room temperature.
The free 6-Aminofluorescein is separated from 6-Aminofluorescein coupled amphiphilic fullerene monomer (AF-1) by size exclusion chromatography on Cross-linked dextran G 75 (HY-148139) column.
The fractions are analyzed by fluorometry for 6-Aminofluorescein emission at 520 nm.

Localization of 6-Aminofluorescein conjugated AF-1 using Fluorescence Microscopy
Human Coronary Artery Endothelial Cells are grown in 8-chamber tissue culture slides.
Exposed to 6-Aminofluorescein-buckysomes for 18 hrs at 37°C, 5%CO2.
Washes with Dulbecco's phosphate buffered saline.
Cells are fixed in 4% Paraformaldehyde for 20 min, and washed twice with Dulbecco's phosphate buffered saline.
Chambers are removed and slides are dried.
Fixed cells are mounted in antifade reagent with DAPI.
Observe using a fluorescence microscope.