Rabbit anti chmp4b

The following primary antibodies were used for Western blotting and immunofluorescence and were kindly provided: mouse anti-gD (LP14) and mouse anti-gB (R69), from Tony Minson (University of Cambridge); mouse anti-VP16 (LP1), from Colin Crump (University of Cambridge); mouse anti-gE, from David Johnson (Oregon Health and Science University); and mouse anti-nectin1 (CK6), from Claude Krummenacher (Rowan University, NJ). Our rabbit VP22-specific antibody (AGV031) was described previously (68 (link)). Commercially available antibodies used in this study included mouse anti-V5, mouse anti-gC, mouse anti-β-actin, mouse anti-CD63, rabbit anti-giantin, mouse anti-CHMP4A, and rabbit anti-CHMP4B (all from Abcam); mouse anti-CD71 (Santa Cruz); and mouse anti-γ-tubulin and mouse anti-α-tubulin (Sigma). Goat anti-mouse IRDye 680RD and goat anti-rabbit IRDye 800CW (Li-Cor Biosciences) secondary antibodies were used as appropriate for Western blotting, while Alexa Fluor-conjugated secondary antibodies (Invitrogen) were used for immunofluorescence. Endocytic structures were labeled by incubating cells with Texas Red-conjugated transferrin (Invitrogen) or Alexa Fluor 488-conjugated transferrin (Molecular Probes) at a concentration of 1 μg/ml for 30 min. Brefeldin A (Sigma) was used at a concentration of 1 μg/ml.

Antibodies used for immunoprecipitation and immunolabeling include rabbit anti-BIN1 exon 13 (custom made, generous gift from Sarcotein), mouse anti-BIN1 exon 17 (clone 99D, Sigma), recombinant monoclonal anti-BIN1 exon 13 (generous gift from Sarcotein), chicken anti-GFP (Abcam), mouse anti-Flag (Sigma), rabbit anti-CHMP4B (Abcam), rabbit anti-V5 (Sigma), rabbit anti-actin (Sigma), mouse and rabbit anti-GST (Santa Cruz), mouse anti-CD63 (Thermo Scientifics), fluorescein-conjugated anti-PI(4,5)P2 IgM (Echelon Bioscience), and fluorescein-conjugated anti-PI(3,4,5)P3 IgM (Echelon Bioscience). Alexa 647-conjugated WGA, Alexa 488-conjugated annexin V, Alexa 555-conjugated annexin V, Alexa 647-conjugated annexin V, and Alexa 647-conjugated phalloidin were purchased from Life Technologies. For imaging and flow cytometry, recombinant anti-BIN1 exon 13 was conjugated with Alexa 647 using a monoclonal antibody labeling kit (Life Technologies).