immunoblots help help help - (Oct/14/2006 )
i am currently reading a paper on the degredation of CFTR by the ubiquitin-proteosome pathway and i am trying to grasp the reason for some of the methodologies. in particular is an immunoblot of CFTR with antibodies towards it. now the protein is separated into detergent soluble and insoluble material. why is this? is that to serve as some sort of control??? HELP
CFTR is linked to actin cytoskeleton which facilitates activation by protein kinase A; as CFTR-actin interaction f.i. by degradation of CFTR or re-constitution to membrane, appears to be dynamic, your observations are highly interesting and should be worked out; take unseparated lysat as control to show total CFTR amount
thanks for the reply kosmodrom. a little bit more elaboration though to my question. i was not conducting an experiment, i was rather try to understand the methodology in the paper. what the investigator did was to immunoblot the detergent soluble and insoluble fractions of CFTR after centrifugation. as i understand the detergent soluble portion is usually what is in the supernatant and the insoluble fraction in the pellet. what i am trying to figure out is why they would blot for both.
the paper is titled "degredation of CFTR by the ubiquitin-proteosome pathway" by ward et al. all the immunoblots had lanes for soluble and inslouble forms of CFTR. I would really appreciate your reply