Help? Immunoprecipitation

brett brett at BORCIM.WUSTL.EDU
Wed Feb 28 13:02:47 EST 1996


>I am trying to discriminate some protein which have same 
>molecular weight with immunogloblin heavy chain.
>If anyone have tip that can solve this problem, please 
>tell me.
>Thanks in advance
>Myung Jong Kim
>My e-maol address is
>        pgs at vision.postech.ac.kr

If I understand your problem, IgH is causing a "smearing" of your band?
This is to be expected when you overload the any protein. If you have a
mAb, try titrating down the amount of Ab used. You will find a concentration
which gives you good signal intensity, and virtually eliminates this problem.
Polyclonals are trickier, as decreasing the [Ig] will significantly decrease
signal intensity. Here you may have to  affinity purify your Ab first.



Brett Lindenbach
    
Program in Immunology                              
Washington University - St Louis                  
brett at borcim.wustl.edu                             

"I own my own pet virus. I get to pet and name her." - Cobain




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