To examine hepatic morphology and assess liver fibrosis, liver specimens were fixed 10% neutral buffered formalin, embedded in paraffin, and cut into 4 μm sections. Specimens were deparaffinized, hydrated, and stained as usual method with standard hematoxylin and eosin staining (H&E) and Sirius red staining as previously described52 (link). For immunohistochemistry (IHC), sections were incubated for 10 min in 3% hydrogen peroxide to block endogenous peroxidase. Antigen retrieval was performed by heating in 10 mM sodium citrate buffer (pH 6.0) for 10 min or incubating with 0.2% pepsin for 10 min. Sections were blocked in protein blocking solution (X9090; Dako, Carpinteria, CA, USA) for 30 min and incubated with primary antibodies, active Caspase-3 (AF835; R&D Systems, Minneapolis, MN, USA), F4/80 (ab6640; Abcam, Cambridge, MA, USA), Fpr2 (ab203129; Abcam), CD68 (ab125212;Abcam) or non-immune sera to demonstrate staining specificity at 4 °C overnight. Polymer horseradish peroxidase (HRP) anti-rabbit (K4003; Dako) or HRP anti-rat IgG (A110-105P; BETHYL, Montgomery, Texas, USA) was used as the secondary antibody. 3,3'-Diaminobenzidine (DAB) (K3466; Dako) was employed for the detection procedure.
Ab203129
Manufactured by Abcam
Sourced in United States, United Kingdom
Ab203129 is a laboratory product from Abcam. It is a piece of equipment used in scientific research and experiments. The core function of this product is to assist with specific laboratory tasks, but a detailed description cannot be provided while maintaining an unbiased and factual approach.
Automatically generated - may contain errors
Lab products found in correlation
K3466, by Agilent Technologies (1 mentions)
Ab6640, by Abcam (1 mentions)
K4003, by Agilent Technologies (1 mentions)
Ab125212, by Abcam (1 mentions)
X9090, by Agilent Technologies (1 mentions)
Af835, by R&D Systems (1 mentions)
Chemidoc xrs imaging system, by Bio-Rad (1 mentions)
Enhanced chemiluminescence, by Merck Group (1 mentions)
Ab7090, by Abcam (1 mentions)
Ab179467, by Abcam (1 mentions)
Ab214486, by Abcam (1 mentions)
Ampkα1, by Cell Signaling Technology (1 mentions)
2 protocols using ab203129
1
Hepatic Morphology and Fibrosis Assessment
2
Western Blot Analysis of Nerve Tissue
Check if the same lab product or an alternative is used in the 5 most similar protocols
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Protein samples were extracted from nerve tissue block and the SCs using RIPA solution. Equal amounts of protein from each treatment were loaded to 10% of sodium dodecyl sulfate-polyacrylamide gel electrophoresis and transferred to polyvinylidene difluoride membranes. The membranes were incubated with Anti-Annexin A1 (ab214486, 1:1000, Abcam, Cambridge, UK), anti-FPR2 (ab203129, 1:1000, Abcam, Cambridge, UK), phospho-AMPKα (2531, Cell Signaling Technology, USA), AMPKα1 (2795, Cell Signaling Technology, USA), and β-actin (ab179467, 1:1000, Abcam, Cambridge, UK) primary antibodies at 4°C overnight. The membranes were further incubated with IgG-horseradish peroxidase goat anti-rabbit secondary antibody (ab7090: 1:2000, Abcam, Cambridge, UK) for 1 hours at room temperature. Signals were developed by enhanced chemiluminescence (Sigma-Aldrich). The stained protein bands were visualized using a Bio-Rad ChemiDoc XRS imaging system and analyzed using Quantity One software.
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