Anti v5 46 0705

Western blotting of cell lysates was conducted as described (22 (link)). Blotting membranes were serially incubated with primary antibodies and species-specific horseradish peroxidase (HRP)-conjugated secondary antibodies (Jackson ImmunoResearch Labs). Immunoreactivity was visualized using Western Lightning® Plus-ECL, Enhanced Chemiluminescence Substrate (Perkin Elmer, Waltham, MA). The primary antibodies used were as follows: Anti-V5 (46-0705, Life Technologies), anti-phospho-c-Jun N-terminal kinase (JNK) (4668, Cell Signaling Technology, Danvers, MA), and anti-total JNK (9252, Cell Signaling Technology). Densitometric analysis of immunoreactive bands was conducted using Multigauge software v. 3.3 (Fuji Film, Tokyo, Japan).

Gels were Coomassie stained and scanned as described (Antrobus and Borner, 2011 (link)). Western blotting was performed as in Borner et al. (2012 (link)). Immunofluorescence microscopy was performed as described for Dmel2 cells (Hirst et al., 2009 (link)). The following antibodies were used: dAP1G (Hirst et al., 2009 (link)), dCLC (Heerssen et al., 2008 (link)), and anti-V5 (46-0705; Life Technologies). Images were adjusted for brightness and contrast in Photoshop (Adobe Systems Europe, Maidenhead, UK) or PowerPoint (Microsoft).