Need help with fluorochrome labelling of antibodies

Dean Hewish deanh at tigger.mel.dbe.csiro.au
Mon Dec 5 22:08:27 EST 1994


I have routinely used the first of the two protocols for some time and
have not had any problems with it. I have labelled everything from
antibodies to peptides using it and it seems ot be reliable enough. It
certainly is very easy to carry out. Mostly I separate unbound
Fluorescein by dialysis rather than a column, because I often don't
have enough protein to afford losses. I have used the conjugates for
FACS and they have worked OK.
I don't know of any commercial labelling kits. The protocols are so
simple and the reagents so cheap that kits would be of dubious value
for money.
I have labelled proteins with rhodamine in the past-that is fairly easy
but I didn't do a lot of testing of the conjugates. Texas red comes as
an isothiocyanate derivative that should handle just like FITC. Other
common fluorochromes like phycoerythrin & allophycocyanin are large
proteins and I do not have any detailed protocols for conjugating them.

Dean R. Hewish
Cell biologist and Flow Cytometrist
CSIRO Division of Biomolecular Engineering
Parkville, Victoria, Australia



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