Antibody to Fluoro-Gold

Each vial of the Antibody to Fluoro-Gold contains approximately 100µl of solution.

Use of the Antibody
Fluoro-Gold can be injected using several different methods, including pressure, iontophoretic and other applications developed by a variety of researchers. See Schmued and Fallon, FluoroGold: “A fluorescent retrograde axonal tracer with numerous unique properties,” Brain Research, 377 (1986) 147-154 as well as Pieribone and Aston-Jones, “The Iontophoretic Application of Fluoro-Gold for the study of afferents to deep brain nuclei,” Brain Research, 475 (1988) 259-271. Many researchers have developed their own modified procedures. Use of the antibody should not be dependent upon the methodology used to employ Fluoro-Gold.
After the Fluoro-Gold has been injected, floating sections (we used thirty um sections from a rat perfused with 4% formaldehyde) are incubated with the Fluoro-Gold Antibody solution overnight at 4 degrees C. Sections are washed, then incubated in Biotinylated GAR (Vector Labs) at 1/1000 for 1 hour at room temperature and washed again. Sections are then incubated in Avidin/Biotin (Vector Labs) at 1/1000 for 1 hour, washed and transferred to Diaminobenzidine (.04%) and Nickel Chloride (2.5%) in 0.1 M NaAcetate with 0.06% H2O2 for six minutes. Sections are then washed, mounted, dried, dehydrated and cover slipped.
It has been our experience that if stored and prepared in the manner set out above, each vial of the antibody should treat 300 to 1000 thirty um sections of the albino rat brain (or similar sized animals) if you use the Vector elite kit. However, you should experiment with the concentration and procedures to determine which best fits your circumstance.

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