How is the cell extract prepared?
We recommend the Dignam method. However, since the Dignam method requires a large amount of cells, we recommend a method of solubilizing with a surfactant such as NP-40 when using a small amount. Please refer to the protocols 401 and 402 from the protocol page. If the above is difficult, you can also use commercially available kits (ProteoExtract Subcellular Proteome Extraction Kit (MERCK MILLIPORE), CelLytic M (SIGMA), etc.). However, if the detergent concentration is 1% or higher, it may prevent the ligand from binding to the target protein during affinity purification, so reduce the detergent concentration to 0.1% by dialysis or dilution before use.