No Protein Induction
Posted 02 March 2005 - 06:02 AM
Posted 02 March 2005 - 08:52 PM
If you are trying to express as a soluble then, the protein might have aggregated into inclusion bodies. first chack if atall the protein is expressed. Run the cell extract/homogenate of induced and uninduced clones on SDS PAGE and check for expression. If you find the band then, but no activity in the relevant sample, then the protein is aggregating into inclusion bodies.
You can try to dissolve the inlcusion bodies with urea or guanidium.HCL and reanture it. During reanturation, inlcude different types ofdetergents and check for stability. Some kits for solubilization and stabilization are available from HAMTON RESEARCH. These are meant for use during crystallization; however, the readymade buffers and solutions give preliminary conditions to start with.
Posted 03 March 2005 - 05:43 AM
Posted 03 March 2005 - 06:04 AM
as you said your protein is a membrane protein, do you extract your protein well (if supposing there's a expression)?