Agent to punch hole in cell membranes - (Feb/06/2008 )
I am trying to use a probe method that binds to hydrophobic pockets of proteins, therefore the native conformation of my protein is crucial. My protein is a golgi transmembrane protein which makes isolation of my protein difficult.
Detergent cant be used because it will bind to my protein and inhibit the probe binding.
I was hoping that I could use density centrifugation to isolate just the golgi, and then use some agent to poke holes in the golgi to allow my probe to enter....bind its heart out all over my protein (Cross link it by UV --known process)....then use detergent to isolate these proteins (now attached to my probe) in an extract so I can run on a gel etc etc.
Does anyone know of an agent that might punch holes in the membrane which will be suitable for my probe to penetrate in and do its job???
There are agents that are used for drug delivery but I need advice.
saponin is a mild detergent which might be useful for your experiment. But definitely try out others detergents like triton (in different conc.) to find the best results.