Rabbit anti glur1

The hippocampal tissues were lysed in lysis buffer (Beyotime Institute of Biotechnology, Haimen, China). After centrifugation at 13,800 × g for 5 minutes at 4°C, the supernatant was collected. Protein samples were separated by 8% or 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis. After being transferred onto a polyvinylidene fluoride membrane (0.45 μm; Millipore, Billerica, MA, USA), the samples were blocked with 5% non-fat dry milk for 1 hour at room temperature and then incubated at 4°C overnight with the following primary antibodies: rabbit anti-GluR1 (1:2,000; Cell Signaling Technology), rabbit anti-GluR2 (1:500; Cell Signaling Technology), and mouse anti-β-actin (1:3,000; Santa Cruz Biotechnology, Santa Cruz, CA, USA). After being rinsed with Tris-buffered saline plus Tween 20 (0.1% Tween 20) three times, the membrane was incubated with secondary antibody against rabbit or mouse IgG conjugated to horseradish peroxidase (1:4,000; Cell Signaling Technology) for 1 hour at room temperature, and treated with enhanced chemiluminescence reagent (Beyotime Institute of Biotechnology). Blots were imaged using X-ray film (Eastman Kodak Co., USA), and images were captured using a scanner (Canon, Japan). The optical density values of the target bands were quantified relative to the level of β-actin using Gel-Pro analyzer 4.0 software (Media Cybernetics, USA).