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Materials
17.3 NaHCO3
8.6g Na2CO3 per 1 liter
Methods
- Antibody (Ab) at 1.5 mg/ml
- Dialyze against Bicarbonate/Carbonate Buffer
- Dissolve FITC fluorescein isothiocyanate in DMSO in a ratio of l mg
FITC/ml DMSO
- Put dialyzed Ab in a small beaker containing a small stirring bar.
- Add directly, very slowly, drop-by-drop using a Pipetteman:
- 10-40 µg FITC/mg Ab (20- 30 µg/mg Ab seems to work best
for goat anti-Mouse Ig)
- Cover and stir for 2 hours at RT.
- Dialyze against PBS - 3 changes over 2 days or run over a G25 fine
column.
- Filter sterilize and store covered at 4° C.
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