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I am about to collaborate on some experiments with a new investigator, and
after some lengthy discussions it appears that the only suitable antibodies
for our experiments are only available in un-conjugated form.
Since I loathe the idea of doing 2 step labelling - with all it's washing
steps (it will be a 2 color experiment so washing/blocking will be painful)
- so I have hit upon the idea of conjugating the antibodies myself. Is this
foolish ?
I have never conjugated an antibody before and would like some input on:-
a. Is this a good idea to attempt this?
b. What is my best chance of sucess, FITC, RPE, biotin?
c. Can someone suggest a protocol which is reliable ?
d. A co-worker tells horror stories of antibodies which conjugate well, and
degrade after 3-4 weeks, is this caused by technique or fate ?
Any suggestions are, as always, appreciated
Peter Chapple
Senior Scientist
Peter MacCallum Cancer Institute
Melbourne AUSTRALIA
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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
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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