?good source for DEAE-dextran?
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,
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
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.
More information about the Methods