How are cell and tissue samples prepared for assays if it is not described in the data sheet?
A.The following general guidelines can be followed: For cell samples: Start with ~2X106 cells, suspend the cell pellet 500 μl (or ~4 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Load the supernatant unto a 10kda spin column for deproteinzation (if indicated in the data sheet-not for enzyme assays). Use the eluate for your subsequent assays. Appropriate dilutions of the sample must be tested in order ensure the readings will fall within the linear range of the standard curve. For tissue samples: Start with 10-100 mg of the tissue, add 500-1000 μl (or ~4-6 volumes) of the assay buffer on ice, homogenize using a Dounce homogenizer (10-50 passes) on ice, until efficient lysis is confirmed, by viewing the cells under the microscope. Spin down the sample and collect the supernatant. Load the supernatant unto a 10
