Which cell permeabilization method is most suitable for this antibody?
Permeabilization should only be required for intracellular epitopes when the antibody requires access to the inside of the cell to detect the protein. However, it will also be required for detection of transmembrane proteins if the epitope is in the cytoplasmic region. Solvents such as acetone and methanol are suitable for permeabilization when detecting intracellular protein. As they disrupt membrane proteins, they are not suitable for use when detecting membrane proteins. Triton or NP-40 will also partially dissolve the nuclear membrane and are therefore very suitable for nuclear antigen staining. However, they are harsh detergents which can disrupt membrane proteins, especially if left on for too long and are not suitable for membrane proteins. Tween 20, Saponin, Digitonin and Leucoperm are much milder membrane solubilizers. They will create pores large enough for antibodies to go through without dissolving the plasma membrane. They are suitable for antigens in the cytoplasm or the
