Lipid patterns of embryonal carcinoma cell lines and their derivatives: Changes with differentiation

Cryoprecipitation of an anti-Pr2 monoclonal IgM cold agglutinin in the presence of GM3 ganglioside.

“…The presence of ganglioside in Mou cryoglobulin was first studied by ELISA: the wells were directly coated by incubation for I h at 37°C, then for 2 h at 4°C, with concentrations of Mou cryoglobulin in GVB2+ ranging from 0.1 to 100 Ag/ml. After being washed as described above, the plates were incubated with a rabbit anti-GM3 serum, raised as previously described (18), and revealed with goat anti-rabbit Ig antibodies labeled with fl-galactosidase (a gift of J. L. Guesdon, Institut Pasteur) using 4 methyl-umbelliferyl-,d-galactopyranoside (Sigma Chemical Co.) as the fluorogenic substrate, as described by Labrousse et al (19). In other experiments, gangliosides were extracted from Mou cryoglobulin and Mou SN IgM, as previously described (20) by a modification of the technique of Cham and Knowles (21).…”