In-Cell Western™ Assay Cell Fixation/Permeabilisation Methods

To enable analysis of cellular proteins for the In-Cell Western (ICW) assay, cells must be immobilized onto a solid microplate surface, and antigens must be made accessible to their antibodies. In addition, permeabilisation is typically required to provide access to the antigen and subcellular localization. Here, two methods are presented for cell fixation and permeabilisation in ICW assays, one using methanol and another using formaldehyde/triton X-100. The methods are compared, and alternate methods are also suggested.