Rabbit anti glua1 antibody

A rat was deeply anesthetized with pentobarbital and transcardially perfused with heparinized saline and subsequently with ice-cold 4% paraformaldehyde in 1 × PBS (pH 7.4). The brain was removed and post-fixed in 4% paraformaldehyde overnight at 4°C, followed by dehydration with 30% sucrose in 1 × PBS. Tissues were sectioned into 30-μm thick coronal sections with a cryostat at −20°C. Sections were blocked with 5% normal goat serum in PBS containing 0.3% Triton X-100 and incubated overnight with primary antibodies: rabbit anti-GluA1 antibody, 1:500 dilution (Millipore, Cat#05–855R), mouse anti-GluA2 antibody, 1:500 (Millipore, Cat# MAB397). Sections were then rinsed and incubated with the goat anti-rabbit or goat anti-mouse Alexa Fluor-conjugated secondary antibody, 1:500 (Molecular Probes Cat# A-11008 and RRID:AB_141371, Cat# A-11004), and were mounted on slides, dried and coverslipped with ProLong Gold anti-fade reagent for staining with the fluorescent reporter 4′,6-diamidino-2-phenylindole (DAPI, Invitrogen). The stained sections were examined with an Olympus BX51 fluorescence microscope.

DIV 14–16 neurons plated on #1.5 glass coverslips were
transferred to ACSF with 1 mM Mg²⁺ for 30 min. Cells were
transferred to ACSF for 30 min then rabbit anti-GluA1 antibody (Millipore
1:250) was live-fed for 15 min at 37°C before being washed in
ice-cold ACSF 2× and fixed in 4% PFA. Cells were then processed for
STED imaging by labeling with mouse anti-PSD-95 primary antibodies and
fluorescent secondary antibody conjugates as described below.