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

******************************************************************************
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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