Immunohistochemistry staining was carried out according to the method previously described [11 (link)]. The brains were fixed in 4% PFA solution for 48 h and were then cut into serial sections (5 μm). The paraffin-embedded hippocampal sections were processed as free-floating slices, including deparaffinized, rehydrated, and pretreated with hydrogen peroxidase. Antigen retrieval was performed by heating in 0.01 mmol/L citrate buffer (PH = 7.2) for 15 min. Slices were incubated with primary antibodies (GLAST, Cell signaling, #5684, 1:50; GLT-1, Abcam, #69098, 1:300) after blocking in the antisera. After incubation with the secondary antibody, sections were placed in DAB reagent (#00–2014, Invitrogen, USA) for 5–10 min at room temperature. After a further rinsing in 0.1 mol PBS, sections were restained with hematoxylin, and were mounted on gelatine-coated slides for observation. The images of the positively stained expressions in the CA1, CA3 and DG regions of the hippocampus were captured at 200 x magnifications by an Olympus BX 41 microscope.
Glast
Manufactured by Cell Signaling Technology
GLAST is a laboratory protein analysis tool used to detect and quantify the expression of the GLAST (Glutamate Aspartate Transporter) protein in biological samples. It is a reliable and sensitive assay that allows researchers to study the role of GLAST in various cellular processes and disease states.
Automatically generated - may contain errors
Lab products found in correlation
Glt 1, by Abcam (1 mentions)
Dab reagent, by Thermo Fisher Scientific (1 mentions)
Bx41 microscope, by Olympus (1 mentions)
Hdac6, by Cell Signaling Technology (1 mentions)
Hdac3, by Cell Signaling Technology (1 mentions)
Hdac5, by Cell Signaling Technology (1 mentions)
P glua1, by Cell Signaling Technology (1 mentions)
Glt 1, by Cell Signaling Technology (1 mentions)
Glun2b, by Cell Signaling Technology (1 mentions)
Glua1, by Cell Signaling Technology (1 mentions)
Hdac1, by Cell Signaling Technology (1 mentions)
Hdac2, by Cell Signaling Technology (1 mentions)
Hdac4, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Enhanced chemi luminescence (ecl), by Bio-Rad (1 mentions)
2 protocols using glast
1
Immunohistochemical Analysis of Hippocampal Astrocytes
2
Western Blot Analysis of Epigenetic and Synaptic Proteins
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Tissue samples were made into protein samples by RIPA lysis buffer and prepared for protein electrophoresis. After electrophoresis, the gels were transferred to polyvinylidene fluoride (PVDF) membranes, then incubated with the indicated antibodies with gentle shaking overnight at 4 °C. The antibodies used were HDAC1 (1:1,000, Cell Signaling Technology, #34589), HDAC2 (1:1,000, Cell Signaling Technology, #57156), HDAC3 (1:1,000, Cell Signaling Technology, #85057), HDAC4 (1:1,000, Cell Signaling Technology, #15164), HDAC5 (1:1,000, Cell Signaling Technology, #20458), HDAC6 (1:1,000, Cell Signaling Technology, #7558), GLT-1 (1:2,000, Cell Signaling Technology, #3838), GLAST (1:2,000, Cell Signaling Technology, #5684), GluN2B (1:1,000, Cell Signaling Technology, #14544), p-GluN2B (1:1,000, Cell Signaling Technology, #5355), GluA1 (1:1,000, Cell Signaling Technology, #13185), p-GluA1 (1:1,000, Cell Signaling Technology, 75574), and β-actin (1:2,000, Cell Signaling Technology, #4967). Next, the PVDF membranes were incubated with specific HRP-conjugated secondary antibodies, and the protein bands were visualized using enhanced chemiluminescence (ECL, Bio-Rad, California, USA). ImageJ software (National Institutes of Health, USA) was employed to quantify the band densities.
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