Figure 1. Pattern of seroreactivities of melanoma patients, healthy controls and patients with other diseases against the melanoma cell line M-NRT.

Total protein extract of the tumor cells were separated by SDS-PAGE, blotted onto nitrocellulose and probed with the sera of the melanoma patients (Panels A–C), of healthy control donors (Panel D, sera 102–110) or patients with cutaneous lymphoma (Panel E, sera 96–98), pancreas carcinoma (Panel E, serum 101) or visceral leishmaniasis (Panel E, sera 111–113). The numbers atop of each lane represent the number of the sera and are used throughout this report. For comparison, melanoma serum 7 was included in all blots. The Western blot analyses were done with the sera at a dilution of 1/6 in a multiple channel blotting/Western blot developing chamber for panels A–E. The letters underneath the lanes indicated the combinations of sera for the multiple probing of the 2D Western blots shown in Figure 2 and summarized in Table 1. Panel F: High-resolution 1D Western blot for comparison of the seroreactivities of melanoma patients and a healthy donor against the melanoma cell line M-NRT. In contrast to the blots shown in panels A–E, the sera were applied 1/200 to isolated lanes from SDS-PAGE in sealed plastic bags which results in a better definition of the bands when compared to blots from multiple blotting devices. Serum 18 is autologous to tumor cell line M-NRT, serum 7 is from a different patient and serum 103 from a healthy donor. The arrows to the left of the lanes indicated antigen bands that are shared between the patients and the healthy controls and occur in every blot. The arrows to the right of the lanes indicated prominent antigen that are detected only by the sera of melanoma patients.