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Antibody against E.coli lysate


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#1 sivakumar

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Posted 27 May 2015 - 08:30 PM

I would like to immunize rabbits with E.coli antigens(french pressed cell lysate) to raise antibodies.

I've heat inactivated lysate at 56 C for 30 min. There are formation of precipitates and if I centrifuge down these precipitates there is a huge difference in protein concentration. How to deal with this.

 

Please suggest me any protocol for heat inactivation of Ecoli lysate.



#2 mdfenko

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Posted 28 May 2015 - 04:44 AM

the precipitate is denatured proteins, hence the difference in protein concentration.

 

do you want to inoculate with specific antigen(s) or are you trying to prepare antibodies to the whole e coli lysate?

 

if to whole lysate then you shouldn't heat the lysate. you lose too many proteins.

 

if for specific antigen(s) then you should ensure that it (they) is (are) still present after heating and clarifying.


Edited by mdfenko, 28 May 2015 - 04:45 AM.

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