Goat anti aqp4

An affinity purified rabbit polyclonal antibody to a predicted antigen (DRTESRQDSLELSS) within ORF2 of AQP4 was prepared by GenScript. The following commercially available antibodies were also used. Mouse anti-Renilla (MBL), mouse anti-HA (Covance), mouse anti-β-actin (Sigma) and goat anti-AQP4 (Santa Cruz).

Confluent astrocyte cultures plated in 100 mm dishes were lysed and equal amounts of total protein were loaded onto 7.5 or 10% SDS-PAGE gels for separation and electrophoretically transferred to nitrocellulose membranes (Whatman, Dassel, Germany). The membranes were probed with polyclonal antibodies to rabbit anti-Cx43, 1:10,000 (Sigma, cat# C6219); mouse anti-GFAP 1: 500 (Sigma cat# G3893), goat anti-GS 1:500 (Santa Cruz, cat# SC-6640), and goat anti-AQP4 1:500 (Santa Cruz, cat# SC-9888), followed by secondary antibody incubation with horseradish peroxidase-conjugated donkey anti-goat IgG, goat anti-rabbit IgG and goat anti-mouse (1:10,000, Santa Cruz Biotechnology, Santa Cruz, CA, United States cat# SC-2020, SC-2004, and SC-2005, respectively). Protein bands were detected using the Immobilon Western Chemiluminescent HRP Substrate (EMD Millipore, MA, United States catalog# WBKLS0100) and images were acquired using the In Vivo FX PRO imaging system (Carestream, Carestream, NY, United States).