Figure 2.
Identification of cellular Trip230 protein by antibodies. Lysates prepared from T24 cells metabolically labeled with [35S]methionine were immunoprecipitated with two antibodies, anti-C5 (lanes 2 to 5) and anti-C5N (lanes 6 to 10), which were raised against two GST-fusion proteins containing amino acids 1,099–1,372 and amino acids 78–376 of Trip230, respectively. These two antibodies recognized a protein of 230 kDa that is not detected by the preimmune serum (PI). Inclusion of the anti-C5 antibody with the GST-C5 fusion protein that contains the corresponding antigen, but not GST alone, resulted in the failure of the antibody to detect the 230-kDa protein, indicating that the antibody is specific. Moreover, results from double immunoprecipitation experiments (∗, lanes 5 and 8) showed the detection of only the 230-kDa protein by both antibodies.