Brain protein lysates were extracted as previously described (11 (link)). Proteins were then loaded in 8–16% agarose precast gels (Biorad) and electroblotted onto 0.45 μm Immobilon PVDF membranes. Membranes were immunoblotted with primary antibodies anti-Iba-1 (rabbit, 1:1,000; WAKO Chemical 019-19741), followed by the appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies and revealed by Clarity western (ECL) substrate (biorad). Quantification was done by determining integrative density of the bands using Thermo Scientific Pierce my Image Analysis Software v2.0. Optical values were normalized over actin.
Pierce my image analysis software v2
Manufactured by Thermo Fisher Scientific
Pierce my Image Analysis Software v2.0 is a computer software solution designed for the analysis and processing of digital images. The software provides core functionalities for image manipulation, measurement, and data extraction. Its primary purpose is to facilitate the study and interpretation of visual information within a scientific or research-oriented context.
Automatically generated - may contain errors
2 protocols using pierce my image analysis software v2
1
Western Blot Analysis of Iba-1 Protein
2
Hippocampus and Cortex Protein Analysis
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Hippocampus and cortex brain proteins were lysates as previously described (27) . Proteins were then loaded in 4-15% agarose precast gels (Biorad) and electroblotted onto 0.45 µm Immobilon PVDF membranes. Membranes were immunoblotted with various primary antibodies, as described in Table 1, followed by the appropriate horseradish peroxidase (HRP)-conjugated secondary antibodies and revealed by enhanced chemiluminescence plus (ECL) solution (GE Healthcare Life Sciences). Quantification was done by determining the integrative density of the bands using Thermo Scientific Pierce myImage Analysis Software v2.0. Optical values were normalized over actin.
Table 1 Represents antibodies used for immunoblot analyses and all related information, including the name of the company, molecular weight, species, secondary antibodies, and dilution rates. Open field was performed to evaluate anxiety-like behaviors, exploration habits, and also locomotor activity as described by Hui et al. (28) . Each mouse was individually recorded and analyzed by the ANY-maze system.
Table 1 Represents antibodies used for immunoblot analyses and all related information, including the name of the company, molecular weight, species, secondary antibodies, and dilution rates. Open field was performed to evaluate anxiety-like behaviors, exploration habits, and also locomotor activity as described by Hui et al. (28) . Each mouse was individually recorded and analyzed by the ANY-maze system.
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