western blot control

Fergus McKenzie mckenzie at naxos.unice.fr
Fri Jun 16 09:38:41 EST 1995


In article <3rnj5g$i5f at neuro.usc.edu> Fiberman, william at neuro.usc.edu
writes:
>I am doing some semi-quantitative westerns to detect changes in protein
>levels.  I need a control antibody that detect a "house-keeping" gene
>that usually does not change in expression level.  Can anyone recommend
>one?

>
Dear Fiberman,
        can you give some more info? I imagine that you would like
a 'housekeeping' protein with a molecular mass significantly
different from your protein of interest. Secondly do you want your
'standard' to be a membrane protein, cytosolic, nuclear or all three?
(we are talking mammalian cells here)

        In an attempt to provoke a discussion I would suggest;
       
         membrane protein- what about a G-protein, such as Gi2 alpha?
        cytosolic protein- MAP kinase (42 or 44 kDa forms).

        In addition, both of the above are ubiquitous, well expressed
and antibodies are widely available. 
(Admittedly you can alter the G-protein expression level at the plasma
 membrane by receptor activation)
        Just a thought!
        Fergus.



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