Scrib-specific antiserum and commercial antibodies
The Scrib-specific polyclonal antiserum Scrib-472 was prepared by Eurogentec by immunization of rabbits with a keyhole limpet hemocyanin (KLH) coupled peptide 1612CSSRRPVRPGRRGLGPVPS1630 (19 C-terminal AA of human Scrib). For the detection of endogenous LPP in MDCKII cells, a LPP-specific monoclonal antibody was used (BD Biosciences, Transduction Laboratories). For the detection of GAL4-fusion proteins in immunocytochemistry, a rabbit polyclonal anti-GAL4 DNA-binding domain antibody (Tebu Bio) was used. Vinculin was detected in cells with a monoclonal anti-vinculin antibody (Sigma, clone hVIN-1), Xpress-tagged proteins with a monoclonal anti-Xpress antibody (Life Technologies). Fluorescently-tagged Alexa-antibodies (Molecular Probes) were used as secondary antibodies for immunofluorescence detection.