mcab/fluorescent conjugation options?

[Jorg Kirberg]

In article <jel-18-1506971454180001 at frans.glaxo.med.ualberta.ca>,
jel-18 at bones.biochem.ualberta.ca (mike craig) wrote:

> Hi,
>  I was wondering if anyone had suggestions for the gentlest possible way
> to conjugate monoclonal antibodies to fluorescent tags? We are having
> difficulty with our standard Fluoroscein isothiocyanate technique, as it
> appears to be causing one antibody to lose it's specificity. Any help
> would be appreciated.
> thanx

Dear Mike,
coming across the same problem I have been using fluorescein-succinimidyl
esters. Boehringer sells some stuff called FLUOS. This however had the
disadvantage of giving somewhat more signal at longer wavelength
(when analysing FLUOS / Pe stained cells on a cytometer more
Pe - %FITC compensation was required as compared to real FITC / Pe stained
cells) - but one could live with that.
Molecular Probes has got some derivatives that have longer spacers
to avaoid this problem but I have not tried it yet.

jorg

(no affiliation with those companies)
---------------------------------------------------------------------

Joerg Kirberg                              Tel. 0031 - 20 - 512 19 98
The Netherlands Cancer Institute           FAX  0031 - 20 - 512 20 11
Div. Molecular Genetics (H4)               E_mail kirberg at nki.nl
Plesmanlaan 121
NL - 1066 CX Amsterdam
The Netherlands