Immunoblot analysis of GST-ALAD fusion proteins.

(A) Immunoblot analysis with MAb anti-GST. The molecular size of the wild type and mutants were 62 kd, except for delTC, which was 58 kd, indicating that ALAD expressed by delTC is shorter than the wild-type ALAD by about 4 kd. (B) Immunoblot analysis with rabbit IgG against antihuman ALAD. Essentially similar results are seen as in panel A, except for the presence of an additional band at about 64 kd, including delTC. This finding suggests that the additional band is likely due to a nonspecific binding of a bacterial protein with IgG. (C) Immunoblot analysis with mouse MAb4. Results are essentially similar to row A. (D) Immunoblot analysis with mouse MAb350. Results are essentially similar to row A. (E) Immunoblot analysis with mouse MAb330. In contrast to polyclonal IgG, MAb4 and MAb350, MAb330 completely failed to recognize delTC.