can coommassie stained gel be western blot
Posted 26 July 2012 - 06:47 PM
can i do western blooting of my proteins which are already coommassie stained, and if yes than how?
Posted 27 July 2012 - 01:46 AM
Posted 27 July 2012 - 06:30 AM
Posted 27 July 2012 - 11:49 AM
Posted 27 July 2012 - 12:02 PM
Edited by proteaMatt, 27 July 2012 - 12:04 PM.
Posted 30 July 2012 - 07:21 AM
first you have to remove traces of fixative (acetic acid is a somewhat reversible fixative). soak the gel in water.
then, use sds (up to 0.05%) in the transfer buffer to help resolubilize the protein.
it may still be inefficient but you should get some transferred.
a bigger concern will be the stain and how it effects post-transfer processing (some of the stain will be removed from the protein and should pass through the membrane).
genius does what it must
i do what i get paid to do