Fig. 5. Triton X-114 extraction solubilizes glycophosphatidylinositol (GPI)-anchored proteins. (A) Western blot probed with anti-A and anti-B antisera. Lane 1, supernatant from phospholipase C (PLC)-treated pellicle; lane 2, supernatant from PLC-treated pellicle in the presence of 2-nitro-4-carboxyphenyl-N,N-diphenylcarbamate and phenylmethanesulfonyl fluoride; lane 3, aqueous phase of Triton X-114 extraction from phase separation of pellicle proteins; lane 4, proteins from the detergent phase of Triton X-114 extraction and phase separation of pellicle. (B) Anti-cross-reactive-determinant developed electroblot of the same samples as in A. The filled arrow points to surface antigens, and the open arrow to 40-60 kDa proteins. 8 % polyacrylamide gels were used.