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Identifying a protein isoform - (Sep/01/2008 )

On my western we saw an expected band which could be a new protein isoform. To identify we wanted to sequence it. I believe you have to have some kind of clean up procedure once you cut out the band from the gel.

Does anyone know where I could find a protocol for this and give me an idea of how much extra work on the back of a western it would be. This will allow me to determine whether I can do it or leave it for another student to look at

Thanks

-Patrick Coovans-

QUOTE (Patrick Coovans @ Sep 1 2008, 06:27 AM)
On my western we saw an expected band which could be a new protein isoform. To identify we wanted to sequence it. I believe you have to have some kind of clean up procedure once you cut out the band from the gel.

Does anyone know where I could find a protocol for this and give me an idea of how much extra work on the back of a western it would be. This will allow me to determine whether I can do it or leave it for another student to look at

Thanks


are you overexpressing? if not, better use 2D gels; you need about ~50 pmol protein for MS

-The Bearer-