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Are there any Mab conjugation experts out there?
We have an IGM Mab which is inactivated when conjugated with FITC by
normal procedures.
We've tried periodate oxidation and conjugation of the carbohydrates to
fluorescein thiosemicarbozide with limited success. The antibody is not
inactivated but we can only achieve a very low F/P ratio which results in
very dim fluorescence.
The application requires a bright, directly coupled Mab - we cannot use
biotin conjugates.
Has anyone had similar experiences? Any tips for increasing the F/P
ratio with the carbohydrate conjugation? Any references on
conjugation methods that do not effect antibody activity? I've done some
pretty extensive lit. searches without much success.
Any suggestions greatly appreciated.
_______________________________________________
Graham Vesey
Australian Environmental Flow Cytometry Group
School of Biological Sciences, .-.--:_:\
Macquarie University, _/ \
Sydney, : AEFCG |
Australia NSW 2109. \_ /
Tel- 612 850 8150 '-''''\__/
Fax- 612 850 8174 V
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CD-ROM Vol 3 was produced by Monica M. Shively and other staff at the
Purdue University Cytometry Laboratories and distributed free of charge
as an educational service to the cytometry community.
If you have any comments please direct them to
Dr. J. Paul Robinson, Professor & Director,
PUCL, Purdue University, West Lafayette, IN 47907.
Phone: (765)-494-0757;
FAX(765) 494-0517;
Web