Murine MAb IgG purification
#1
Posted 15 March 2002 - 07:01 PM
Thanks. : )
#2
Posted 21 March 2002 - 03:13 AM
Insulin + Transferrin + Ethanolamine + Selenium
in F12:DME medium....
adapt your hybridoma cell over 2 weeks with gradual reducing FBS % to 0.5%....
then make 1000mL of media... incubate until media changes its color to yellowish.... not too yellow I mean..
prep. sup. add... ammonium sulfate to precipitate...
spin down.... resuspend pellet .... dialysis ....
QAE resin for 2hrs to remove Ig from FBS...
sup... filter... concentration... amicon centricon...
SDS-PAGE to check your final protein...
maybe you can do western blot analysis with anti-mouse IgGXX...
Good luck!
Hyun-Seok Kang
Department of Molecular Biology
Cellular Immunology Lab.
samuelmb@chollian.net













