?good source for DEAE-dextran?

Uttam Rajbhandary bhandary at wccf.mit.edu
Wed Dec 1 15:33:00 EST 1993


In article <2d5f39$p5o at oac4.hsc.uth.tmc.edu>, gsbs1044 at UTSPH.SPH.UTH.TMC.EDU writes...
>Does anyone have a favorite supplier of DEAE-dextran?  I want it, as you
>may have guessed, for transfecting COS cells.  I'd be interested to 
>read any tried and true protocols as well.
>thanks in advance,
>tim ritty 


Believe it or not, I've been using "plain ol'" Sigma as the source.  I make a 
2mg/ml stock in pbs, and as a working stock, make 1 vol of the dex stock, 1 
vol 1 M tris HCl pH 7.3 and 8 volumes DME (no serum), final deae-dex of 200 
ug/ml.  
I split a 10 cm confluent cos-1 plate to 10 ml, and inoculate 6 cm dishes with 
0.5 ml of the 10 ml suspension.  48 hours later, I the rinse 2x with dme (no 
serum), then add 5-10 ug of dnas in 2 ml of the working dex/dme solution.  
5hours inc at 37.  Rinse 2x with dme (no serum), and inc with 2 ml of 0.1mM 
chlroquine in DME for 2 hours at 37.  Rinse 2x with dme, then add complete 
media.  Harvest 48-72 hours later depending upon the particular purpose.  
Reproducibility appears quite good.

H. Drabkin



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