Brains were carefully removed and postfixed in 4% paraformaldehyde overnight at 4°C. After dehydration, coronal sections (10 μm) containing the hippocampus were cut using a freezing microtome (CM1950, Leica). The sections were permeabilized with 0.3% Triton X-100 in normal goat serum for 1 hour. Then, the sections were incubated with GFAP antibody (1:400, no. GB11096, Servicebio) overnight at 4°C. After washing, the sections were incubated with Cy3-conjugated goat anti-rabbit secondary antibody (1:300; no. GB21303, Servicebio) for 1 hour at room temperature. DAPI was used to stain nuclei. All sections were mounted onto glass slides with cover glass using 50% glycerinum. For astrocyte morphology analysis, fluorescence images were acquired using a confocal laser-scanning microscope (Nikon ECLIPSE C1). A stack of images of each slice spanning a thickness of 5 μm were taken at an optical step of 1 μm. The freehand selection tool in ImageJ was employed to measure the cell body area. Sholl analysis (starting radius: 4 μm, ending radius: 50 μm, radius step size: 2 μm) was performed to analyze astrocyte branches and processes.
Gb11096
Manufactured by Wuhan Servicebio Technology
Sourced in China
The GB11096 is a laboratory equipment designed for automated and quantitative analysis. It functions as a microplate reader, capable of measuring optical density or fluorescence within microtiter plates. The core purpose of this device is to facilitate various assays and experiments that require precise and reproducible measurements of samples in a high-throughput format.
Automatically generated - may contain errors
Lab products found in correlation
Eclipse c1, by Nikon (3 mentions)
Cm1950, by Leica (1 mentions)
Gb21303, by Wuhan Servicebio Technology (1 mentions)
Gb11027, by Wuhan Servicebio Technology (1 mentions)
Gb11195, by Wuhan Servicebio Technology (1 mentions)
Gb13105, by Wuhan Servicebio Technology (1 mentions)
G1012, by Nikon (1 mentions)
G1012, by Wuhan Servicebio Technology (1 mentions)
Gb11105, by Wuhan Servicebio Technology (1 mentions)
Gb11181, by Wuhan Servicebio Technology (1 mentions)
Gb12139, by Wuhan Servicebio Technology (1 mentions)
Ap0564, by ABclonal (1 mentions)
Gb113373, by Wuhan Servicebio Technology (1 mentions)
Anti neun antibody, by Wuhan Servicebio Technology (1 mentions)
Gb11063, by Wuhan Servicebio Technology (1 mentions)
Gb11119, by Wuhan Servicebio Technology (1 mentions)
Gb11285, by Wuhan Servicebio Technology (1 mentions)
Gb13105 1, by Wuhan Servicebio Technology (1 mentions)
Eclipse e100 microscope, by Nikon (1 mentions)
β actin 3700, by Cell Signaling Technology (1 mentions)
7 protocols using gb11096
1
Quantitative Astrocyte Morphology Analysis
2
Immunohistochemical Analysis of Colon and Brain
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ZO-1 and F4/80 staining of the colon was detected by staining the colonic tissue sections (5 μm) with anti-ZO-1 antibody (Servicebio, GB11195, 1:200 dilution) in PBS, anti-F4/80 antibody (Servicebio, GB11027, 1:1000 dilution) in PBS and goat-anti-rabbit CY3 conjugated antibody (Servicebio, GB21303, 1:300 dilution) in PBS. Finally, the sections were counterstained with DAPI (Servicebio, G1012). At a temperature of − 18°C, 20 μm frozen brain sections were cut using a cryostat. The brain slices were blocked with 3% bovine serum albumin for 30 min at room temperature and then incubated with the primary antibodies at 4°C overnight. The primary antibody anti-Iba1 (Servicebio, GB13105, 1:500 dilution) and anti-GFAP (Servicebio, GB11096, 1:800 dilution) were used. After washing with PBS, the sections were incubated with the secondary antibodies at 37°C for 50 min. The secondary antibody goat-anti-rabbit Cy3 conjugated antibody (Servicebio, GB21303, 1:300 dilution) were used. Finally, the sections were counterstained with DAPI (Servicebio, G1012) and then imaged with microscope (Nikon Eclipse C1). Quantification of positively stained cells in the PFC and HIP regions were used ImageJ.
3
Immunohistochemistry and Immunofluorescence Analysis of Mouse Brain
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In brief, the mouse brain was embedded in paraffin and sliced into coronal sections (5 μm) of striatum and SN. Paraffin sections were deparaffinized, hydrated, antigen retrieved, incubated with H2O2 and blocked with serum.
For immunohistochemistry experiments, coronal sections were incubated with the primary antibody anti‐TH (Servicebio, GB11181) at 4°C overnight and the secondary antibody at room temperature for 2 h. Slides were visualized by diaminobenzidine staining as the chromogen and observed by a light microscopy (Nikon E100, Japan).
For immunofluorescence experiments, coronal sections were incubated with the primary antibody (anti‐IBA‐1; Servicebio, GB11105; anti‐GFAP; Servicebio, GB11096) at 4°C overnight and secondary antibody at room temperature for 1 h. Nuclei was stained by 4′,6‐diamidino‐2‐phenylindole solution. Slides were visualized and observed by a fluorescence microscope (Nikon Eclipse C1, Japan).
For immunohistochemistry experiments, coronal sections were incubated with the primary antibody anti‐TH (Servicebio, GB11181) at 4°C overnight and the secondary antibody at room temperature for 2 h. Slides were visualized by diaminobenzidine staining as the chromogen and observed by a light microscopy (Nikon E100, Japan).
For immunofluorescence experiments, coronal sections were incubated with the primary antibody (anti‐IBA‐1; Servicebio, GB11105; anti‐GFAP; Servicebio, GB11096) at 4°C overnight and secondary antibody at room temperature for 1 h. Nuclei was stained by 4′,6‐diamidino‐2‐phenylindole solution. Slides were visualized and observed by a fluorescence microscope (Nikon Eclipse C1, Japan).
4
Spinal Cord Injury Histology
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Euthanasia of rats by 2% sodium pentobarbital injection followed by intracardiac perfusion fixation with PBS and 4% paraformaldehyde. The spinal cord was embedded in paraffin and then cut into 20 µm thick sections using electric slicer (Leica, Germany). HE and immunofluorescence staining using primary antibody including anti‐iNOS antibody (GB11119, Servicebio, China), anti‐F4/80 antibody (GB113373, Servicebio, China), anti‐ARG1 antibody (GB11285, Servicebio, China), anti‐CD31 antibody (GB11063, Servicebio, China), anti‐Tuj‐1 antibody (GB12139, Servicebio, China), anti‐GFAP antibody (GB11096, Servicebio, China), anti‐GAP43 antibody (GB11095, Servicebio, China), anti‐NG2 antibody (GB115534, Servicebio, China), anti‐TOMM20 antibody (GB111481, Servicebio, China), anti‐p‐P70S6K antibody (AP0564, Abclonal, China), anti‐NeuN antibody (Servicebio, China) in samples were used to evaluate lesion cavity and nerve regeneration.
5
Glial Cell Visualization and Quantification
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Microglia and astrocytes were immunohistochemically labeled with Iba-1 and GFAP respectively [24 (link)]. Paraffin sections were washed with a dewaxing solution, absolute ethanol and ethanol, and then placed in the citric acid antigen repair buffer (pH 6.0) repair box to repair the antigen. They were blocked in 1% normal goat serum for 30 min at room temperature. Immunohistochemical visualization of glial fibrillary acidic protein (GFAP) and Iba-1 was performed with Rabbit anti mouse GFAP antibody (1:1000, Gb11096, Servicebio, Wuhan, China) and Rabbit anti mouse Iba-1 antibody (1:2000, gb13105-1, Servicebio, Wuhan, China). Finally, the slides were dehydrated and sealed with alcohol, absolute ethanol, n-butanol and xylene in turn. A Nikon eclipse E100 microscope (Nikon company in Tokyo, Japan) was used for visualization after drying. Image J analysis software 6.2 (media cybernetics, silver spring, USA) was used for quantification, and the integral optical density (IOD) of positive stained cells in hippocampal CA1 and CA3 regions was analyzed at 40 times magnification.
6
Kaempferol Modulates Neuroinflammation
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Kaempferol was purchased from Chengdu Alfa biotechnology CO., LTD (Chengdu, China). Antibodies against caspase-3 (ab13847), BDNF (ab108319), TrkB (187041), COX-2 (ab15191), CitH3 (ab281584), Galectin-3 (ab76245) and ICAM-1 (ab171123) were purchased from Abcam (Cambridge, UK). Antibodies for β-actin (3700), p-PI3K (4288), PI3K (4257), Bcl-xl (2764T), p-AKT (9271S), AKT (9272S), MyD88 (4283), p-NF-κB (3033), NF-κB (4764), p-JAK1 (3331), JAK1 (88617), p-STAT3 (9145) and STAT3 (9139) were purchased from Cell Signaling Technology (Beverley, CA, USA). Anti-Bax antibody (50500-2-Ig), anti-Bcl-2 antibody (12789-1-AP), anti-ZO-1 antibody (1773-1-AP) and anti-occludin antibody (13409-1-AP) were the products of Proteintech (Wuhan, China). Antibodies for TLR4 (sc-293072) was provided by Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Antibodies against NeuN (GB13138-1), Iba-1 (GB13105-1) and GFAP (GB11096) were provided by Servicebio (Wuhan, China).
7
Penile and MPG Tissue Analysis
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Freshly dissected penile and MPG tissues were harvested and fixed with 4% paraformaldehyde for further histological staining analysis. Masson trichrome staining was first carried out to determine smooth muscle and collagen expression levels in penile tissues. A 5‐μm slice was prepared and stained following the manufacturer's instructions, and the smooth muscle was stained red while the connective tissue appeared blue. For immunofluorescence staining, the penile sections were incubated with primary antibodies including α‐SMA (1:300, Servicebio, GB111364), eNOS (1:500, Abcam, ab66127), nNOS (1:500, Servicebio, GB11145), Caspase‐3 (1:200, Servicebio, GB11009‐1), and the MPG sections were covered by primary antibodies including Tuj1 (1:2500, Servicebio, GB11139), and GFAP (1:2500, Servicebio, GB11096) at 4°C overnight. After rinsing the slices with PBS, secondary antibodies were adopted for 1‐h immersion. Nuclei were stained with DAPI. Images were observed, and the interesting areas were captured under a confocal laser scanning microscope (Zeiss LSM 710) and a fluorescence microscope (NIKON, Ti2). FIJI/ImageJ software (National Institutes of Health) was used to carry out image analysis.
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