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Murine MAb IgG purification


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#1 henri

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Posted 15 March 2002 - 07:01 PM

Is there anyone could give me some suggestions on the purfication stratages for murine monoclonal antibodies , such as IgG1, IgG2a, IgG2b?
Thanks.  : )

#2 TRUONGTHANG

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Posted 21 March 2002 - 03:13 AM

Preparation of ITES serum free medium...
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





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