Homogenization and solubilization

Once an appropriate source has been recognized, the next step is to get the protein in solution. For proteins in biological ?uids, like as cell culture medium or blood serum,  this  is already  the  case,  but  for  several  proteins  the  tissues  and  cells required to be disrupted and broken open lysed.  Additionally  to  the  process  elaborate  there,  another  easy  way  of  breaking open cells which do not have a rigid cell wall to release the cytosolic  contents  is osmotic  lysis.  The water in the surrounding  solution  diffuses  into the more  concentrated  cytosol  causing  the cell to burst and swell when  animal  cells  are placed  in a hypotonic  solution  (like  as water   or   a  buffered   solution   without   added   sucrose). Differential centrifugation is then employed to erase contaminating subcellular organelles.   Those proteins which are bound to membranes need a further solubilization step. Further isolation through other centrifugation the appropriate membrane is treated with a detergent like as Triton X-100 to disrupt the lipid bilayer and to release the integral membrane proteins into solution.