Electroporation of Antibodies

brett at BORCIM.WUSTL.EDU brett at BORCIM.WUSTL.EDU
Tue Mar 28 12:03:06 EST 1995


>In article <3kmvrq$gce at rover.ucs.ualberta.ca>,
>eatkinso at gpu3.srv.ualberta.ca (gettin' better) wrote:
>
>> Hello!
>> In order to appease the powers-that-be at a certain journal, we find it 
> (deleted) 
>> 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,
> I don't know if it would work or not, but have you thought
>of lipofection?  I've done transfections of cos cells with both
>electroporation and lipofection (LipofectAce, Gibco/BRL) and have
>gotten consistently better results with the latter.  Less cell 
>death and better representation of the complex transfected pool 
>of constructs.  If it works for getting ab's into the cells, 
>it might be a better way to go.  Just a thought.
>
>- Dan
>Positional cloning...it's not the kill, it's the thrill of the chase.

Hello, just wanted top add that it is possible to use lipofection to introduce
proteins into cells. I have used the pT7T7 system to co-transfect plasmids
containing genes driven from the T7 Pr and T7 RNA Polymerase. Cells used were
L929 (rather tranfectable), and best results (%trfxn efficiency) were obtained
with lipofectamine. T7RNAP is ~98kDa, and Ab's range from 150kDa to
>900kDa., for what it's worth. Also, you might consider the net charge on
these proteins
for their ability to form transfection complexes. Maybe the T7 is being brought
in courtesy of the plasmid, which acts as a carrier. Finally, do the experiment,
it's the only way to know! ps let us know how it works.



Brett Lindenbach
    
Lucille P. Markey Student in Human Pathobiology      _  
Program in Immunology                               /_\ 
Washington University - St Louis                   |\d/|
brett at borcim.wustl.edu                              \_/ 






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