Electroporation of Antibodies
gettin' better
eatkinso at gpu3.srv.ualberta.ca
Tue Mar 21 11:45:46 EST 1995
Hello!
In order to appease the powers-that-be at a certain journal, we find it
necessary to introduce antibodies into mammalian cells. Apparently this
has been done with fairly high efficiency using electroporation. I've
found SOME references on the subject but haven't really found definitive
information. The most useful reference that I've dug up seems to use
VERY high concentrations of antibody (2mg/ml FINAL concentration, after
adding to cells in the cuvette). Is this necessary? Does anybody out
there have any experience with this kind of thing? We're trying to
interfere with a signaling pathway by interfering with the association
between a src-family tyrosine kinase and another protein. Any ideas
about how long after zapping the cells we should wait for them to recover
before we stimulate them?
Any advice you can give would be GREATLY appreciated! And if you can
think of anything else that we should consider that I haven't brought up
in this post, PLEASE fell free to suggest-away!
Thanks in advance for your help.
eric
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Eric Atkinson * "When they said: 'Repent, repent',
Dept. of Biochemistry * I wondered what they meant".
University of Alberta * Leonard Cohen
Edmonton, Alberta *
Canada *
T6G 2H7 *
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