Supporting information for Screaton et al. (2003) Proc. Natl. Acad. Sci. USA, 10.1073/pnas.0431215100
Supporting Figure 5Fig. 5.
Characterization of the antibodies used in this study. (a) Two of the FADD antibodies used in this study, the monoclonal from Transduction Laboratories (TL) and our polyclonal BUR385, were characterized by Western blotting of two-dimensional gels containing total MCF10a cell lysates. The two spots represent the unphosphorylated (more basic spot) and the phosphorylated (more acidic spot) forms of FADD seen in numerous previous publications on one-dimensional and two-dimensional gels (cf. ref. 1). Our second polyclonal anti-FADD, BUR386, was analyzed by standard Western blotting. (b) Our anti-mouse FADD antibody BUR404 was characterized by Western blotting of total cell lysates from FADD–/– MEFs vs. FADD-rescued MEFs. Note that, despite the presence of a weak background band on the blot, the FADD–/– MEFs showed no immunofluorescent signal above background with this antibody. (c) The MLH1 mAb from Oncogene Research was characterized by Western blotting lysates of an MLH1-deficient cell line, HCT116 (H), and an MLH1-expressing cell line LOVO (L). (d) Characterization of the sheep anti-MBD4 antibody (self-explanatory.)1. Scaffidi, C., Volkland, J., Blomberg, I., Hoffmann, I., Krammer, P. H. & Peter, M. E. (2000) J. Immunol. 164, 1236–1242.