detect splice variants in living cell

[Peter Ellis]

On Sun, 13 Jul 2008, DK wrote:
>>"Zhong Silin" <stxsz1 from nottingham.ac.uk> wrote:
>>
>> How can we detect alternative splicing in a living cell?
>>
>> I assume that one way is to insert a fluorescent/epitope tag to a specific
>> region of that protein, which is present/absent in its splice variants.
>>
>> Any suggestion please?
>
> Why not Northern?

A)  Because he wants to look at protein levels, not RNA levels

B)  Because he wants to do it in living cells


I don't see the necessity for incorporating a reporter tag into the gene - 
would it not be simpler to raise an antibody specifically to the variant 
splice isoform?  Introducing a tag would seem to run a large risk of 
altering the function of the tagged exon.

Peter