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I see that assay buffer can be used to extract cells for use with this kit. But there is not protocol describing how to use the assay buffer to produce the cell extract. Please provide a protocol.
Asked on Apr 09 2014
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. For enzymztic assay kits, it may be useful to deproteinate samples now by loading the supernatant into a 10kda spin column for deproteinzation. (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.
Jeremy KasanovAbcam Scientific Support
Answered on Apr 09 2014