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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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