Figure 5.

Analysis of centrin in Xenopus and starfish eggs by Western blot analysis. Total proteins from low speed Xenopus egg extracts (Left) and from starfish fertilized eggs (Right) were submitted to SDS/PAGE on a 12% polyacrylamide gel. For Xenopus, two different lanes were incubated with 20H5 mAb (lane 1) and anti-Cdc31p Abs (lane 2), using biotin-conjugated anti-mouse and anti-rabbit secondary Abs, respectively. For starfish, the same lane was incubated first with 20H5 mAb (lane 3), and the detection performed as described, and second with rabbit anti-Cdc31p Abs (lane 4), using peroxydase-conjugated anti-rabbit secondary Abs. The slow-migrating band in the 20H5 pattern is specifically recognized by anti-Cdc31p Abs in Xenopus and starfish.