After the remaining mice in each group had been euthanized, the brains were quickly removed from the ice surface, the hippocampus on both sides was separated and placed in lyophilization tubes and then quickly immersed in liquid nitrogen, the tissues were rapidly frozen and then immediately transferred to an −80°C refrigerator for storage, and set aside for subsequent testing. The protein concentration in the supernatant was detected using a bicinchoninic acid assay kit (Beyotime, Shanghai, China) according to the manufacturer’s instructions. Proteins were separated using 10% sodium dodecyl sulfate–polyacrylamide gel electrophoresis. The primary antibodies used were as follows: PDGFR-β (1:1,000, ab32570, Abcam), α-SMA (1:1,000, ab7871, Abcam), occludin (1:1,000, ab236127, Abcam), claudin-5 (1:1,000, ab131259, Abcam), MMP9 (1:1,000, GB11132, Servicebio), β-actin (1:2,000, ab8226, Abcam), and GAPDH (1:2,000, ab8245, Abcam). The membranes were then incubated with horseradish peroxidase-conjugated anti-rabbit or anti-mouse IgG (1:3,000; Cell Signaling Technology) for 1 h at room temperature and visualized by exposure to Kodak film after detection with chemiluminescent reagent (Millipore). The strip density was quantified with ImageJ analysis software.
Ab236127
Manufactured by Abcam
Ab236127 is a laboratory equipment product from Abcam. It is a device designed for a specific function in the laboratory setting. No further details about its intended use or capabilities are provided.
Automatically generated - may contain errors
Lab products found in correlation
Ab32570, by Abcam (3 mentions)
Ab131259, by Abcam (3 mentions)
Ab7871, by Abcam (3 mentions)
Ab8245, by Abcam (3 mentions)
Horseradish peroxidase conjugated anti mouse or anti rabbit igg, by Cell Signaling Technology (2 mentions)
Chemiluminescent reagent, by Merck Group (2 mentions)
Bicinchoninic acid assay kit, by Beyotime (2 mentions)
Gb11132, by Wuhan Servicebio Technology (2 mentions)
Ab8226, by Abcam (2 mentions)
Chemidoc imaging system, by Bio-Rad (1 mentions)
Ab76003, by Abcam (1 mentions)
3 protocols using ab236127
1
Hippocampal Protein Expression Analysis
2
Protein Expression Analysis of Brain Tissue
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Brain tissue ground on ice for 20 min using a glass homogenizer, centrifuged at 12000 rpm at 4°C for 15 min in a low-temperature ultracentrifuge, and the supernatant was carefully aspirated into centrifuge tubes. This was followed by protein concentration determination, protein denaturation, SDS-PAGE electrophoresis, and membrane transfer. The membranes were incubated overnight at 4°C with the following primary antibodies: PDGFR-β (1:1000, Abcam, ab32570), α-SMA (1:1000, Abcam, ab7871), Occludin (1:1000, Abcam, ab236127), Claudin5 (1:1000, Abcam, ab131259), MMP-9 (1:1000, ab76003), and GAPDH (1:2000, Abcam, ab8245). Finally, blot imaging was performed using a ChemiDoc imaging system (Bio-Rad).
3
Hippocampal Protein Extraction and Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
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After the remaining mice in each group had been euthanized, the brains were quickly removed from the ice surface, the hippocampus on both sides was separated and placed in lyophilization tubes and then quickly immersed in liquid nitrogen, the tissues were rapidly frozen and then immediately transferred to an -80ºC refrigerator for storage, and set aside for subsequent testing. The protein concentration in the supernatant was detected using a bicinchoninic acid assay kit (Beyotime, Shanghai, China) according to the manufacturer's instructions. Proteins were separated using 10% sodium dodecyl sulfatepolyacrylamide gel electrophoresis. The primary antibodies used were as follows: PDGFR-β (1:1000, ab32570, Abcam), α-SMA (1:1000, ab7871, Abcam), occludin (1:1000, ab236127, Abcam), claudin-5 (1:1000, ab131259, Abcam), MMP9 (1:1000, GB11132, Servicebio), β-actin (1:2000, ab8226, Abcam), and GAPDH (1:2000, ab8245, Abcam). The membranes were then incubated with horseradish peroxidaseconjugated anti-rabbit or anti-mouse IgG (1:3000; Cell Signaling Technology) for 1 h at room temperature and visualized by exposure to Kodak lm after detection with chemiluminescent reagent (Millipore). The strip density was quanti ed with ImageJ analysis software.
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