Help! Panning

Clifford Beall beall.3 at OSU.EDU
Tue Aug 8 09:11:35 EST 1995


In article <9508080226.AA12954 at gipac.shinshu-u.ac.jp>,
tnakane at GIPAC.SHINSHU-U.AC.JP (tokio nakane) wrote:

> Hello,
> We are currently trying to clone a G protein-coupled receptor by panning
method.
> We constructed a rat brain cDNA library (pME18S vector; SR-alpha promoter)
> and transfected the library to COS-7 cells by DEAE dextran method.  After
> 72 hrs, we could not detach cells from dishes by the treatment with PBS and
> 5 mM EDTA at 37 C for 30 min.  The untransfected cells were easily detached
> by the treatment.  Any advice would be much appreciated.
> 
> Tokio    

Dear Tokio,

You might try 293 cells (human embryonic kidney).  They are generally
easier to detach from plates.

Cliff Beall
Ohio State Neurobiotechnology Center
beall.3 at osu.edu



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