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B Per for total bacterial protein extraction


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#1 JamASharpe

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Posted 20 November 2013 - 04:37 PM

Hi all,

 

I am in need of clarification of something that is concerning me.  I am wanting to extract a transmembrane protein localised in the cytoplasmic membrane of E. coli for western blotting and hence I bought B Per from Pierce Scientific.  It states that it is an easy to use, one reagent method for gentle lysis of bacterial cells to extract both soluble and insoluble proteins and soluble proteins from inclusion bodies.  However, it does not specifically state that it does total protein extraction and hence the transmembrane proteins.

 

Can anyone put my mind at ease that this solution will extract membrane bound proteins, or do I need to write this off as an extravegant and needless purchase which I will need to get another kit that is more suitable for the job.

 

Any help on this matter would be very appreciated.

 

Many thanks.

 



#2 Missle

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Posted 21 November 2013 - 07:23 AM

Hi.  I'm not sure that B-per will extract membrane proteins.  In my experience, it doesn't even do a great job lysing the cells.  BioRad has a large variety of extraction reagents including one that specifically speaks to membrane proteins (including bacterial - I think), ReadyPrep Membrane II.  I've never used it but maybe their manual or technical support can help?



#3 JamASharpe

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Posted 22 November 2013 - 01:31 AM

Thanks for your input Missile.  I may just do an SDS extraction as I don't need to do downstream processing, I just need to check expression. 

 

I'll do both (SDS and B Per) in parallel and see what bites!






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