For immunohistochemical analysis, thin sections blocked by 5% goat serum followed by incubating in specific primary antibodies. PBS was applied to wash the sections three times. Then, samples was stained with horseradish peroxidase-conjugated secondary antibodies and visualized by substrate DAB. Images were taken with a microscope (Leica, 200 ×) under same acquisition settings for each section. The primary antibodies were used as follow: TGF-β (MAB240-100, R&D System, 1:600 dilution), F4/80 (LS-C96373-100, Lifespan, 1:1000 dilution), CD68 (ab125212, Abcam, 1:600 dilution), IL-1β (SRP8033, Sigma, 1:1000 dilution), TNF-α (ab6671, Abcam, 1: 600 dilution).
Ls c96373 100
Manufactured by LifeSpan BioSciences
LS-C96373-100 is a laboratory equipment product. It is a 100-well microplate designed for various biological assays and experiments. The product provides a standardized platform for conducting tests and analyses in a multi-well format.
Automatically generated - may contain errors
4 protocols using ls c96373 100
1
Immunohistochemical Analysis of Inflammatory Markers
2
Immunohistochemical Analysis of Liver Inflammation
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Samples of liver tissues were excised, and fixed in 10% formalin for 24 h at room temperature. Liver tissues were embedded in paraffin, cut into 5 μm thin sections, and subsequently deparaffinization, rehydration, antigen retrieval and 5% normal goat serum blocking for 4 h at room temperature. The slides were incubated in specific primary antibodies for 4 h, and followed by PBS washing for three times. Then, slides were incubated with horseradish peroxidase‐conjugated secondary antibodies for 2 h, and visualized using substrate diaminobenzidine. The following primary antibodies were used: CD68 (ab125212, Abcam, 1:800 dilution), F4/80 (LS‐C96373‐100, Lifespan, 1:800 dilution), IL‐1β (SRP8033, Sigma, 1:800 dilution), TNF‐α (ab6671, Abcam, 1:1000 dilution). One sample from each mouse was detected, and three high‐power fields at 200× were analyzed for each slide using a microscope (Leica). The number of positive cells were counted.
3
Intestinal Inflammation Immunohistochemistry
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Inflammatory infiltration was evaluated through immunohistochemical analysis of intestinal tissues. Sections from each groups were blocked using 5% goat serum and subsequently incubated in specific primary antibodies, including F4/80 (LS-C96373-100, Lifespan, 1:1000 dilution), Ly6g (GTX40912, GeneTex, 1:500 dilution), IL-6 (ab9324, Sigma, 1:1000 dilution), and TNF-α (ab6671, Abcam, 1: 600 dilution). Slides then were washed thrice with PBS and stained with horseradish peroxidase-conjugated secondary antibodies. Substrate DAB was applied for visualization. Images were obtained under high-power fields with a microscope.
4
Immunohistochemistry and Apoptosis Analysis
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Immunohistochemistry was performed as described previously 46 (link),47 (link). The following antibodies were used: F4/80 (LS-C96373-100, Lifespan, 1:500 dilution), CD11b (ab6672, Abcam, 1: 1000 dilution) and α-SMA (ab7817, Abcam, 1: 500 dilution). Immunofluorescence TUNEL assay was used to assess apoptosis. Tissue sections were placed and fixed in 4% paraformaldehyde, and incubated with immunofluorescent TUNEL reaction mixture for 1 h in box. DAPI was used to stain nuclei. Immunofluorescence images were obtained in Olympus IX71 microscope (Tokyo, Japan, Leica).
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