Liver sections were stained with hematoxylin and eosin (H&E) for necrosis evaluation. Sirius Red staining was performed to evaluate fibrosis. Fast green was used for background staining. Immunohistochemical staining (IHC) for collagen I was performed by using anti-collagen I antibody (Millipore), followed by a Broad Spectrum (AEC) Histostain-Plus kit (Invitrogen). No staining was observed in the absence of the primary antibody. For the computer-assisted quantification assessment, the integrated optical density (IOD) was calculated from 10 random fields per section containing similar size portal tracts and central veins at ×100 and using Image-Pro 7.0 Software (Media Cybernetics, Bethesda, MD, USA). The results were expressed as fold-change over the controls.
Anti collagen 1 antibody
Manufactured by Merck Group
Sourced in United States
The Anti-collagen I antibody is a laboratory reagent used in research applications. It is designed to specifically bind and detect the presence of type I collagen, which is a structural protein found in various tissues and organs. The antibody can be utilized in various analytical techniques, such as immunohistochemistry and Western blotting, to investigate the expression and distribution of type I collagen in samples.
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3 protocols using anti collagen 1 antibody
1
Histological Analysis of Liver Fibrosis
2
Isolation and Characterization of Fibrocytes
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Peripheral blood was mixed 1:3 with red blood cell lysis buffer (Solar, China) and incubated at room temperature for 10 min. Each mixture was centrifuged for 10 min at 450 g. The precipitate was collected and washed with red blood cell lysis buffer and PBS. The resulting leukocyte preparations were incubated with antibodies to CD45 and CD34 (both from Biolegend, US). Alternatively, the leukocyte preparations were incubated in fixation/permeabilization solution for 20 min, washed with Perm/Wash buffer (BD Biosciences, US), and incubated with anti-collagen I antibody (Millipore, US) for 30 min. The cells were washed twice and incubated with anti-TSHR antibody (Novus Biologicals, US), aptamer, or the aptamer library. After washing with Perm/Wash buffer, the cells were evaluated by flow cytometry.
To test fibrocytes, the aptamer was denatured at 95°C for 10 min, renatured on ice for 10 min, and incubated with suspensions of fibrocytes for 30 min at room temperature. After washing twice with DPBS, the fibrocytes were evaluated by flow cytometry.
To test fibrocytes, the aptamer was denatured at 95°C for 10 min, renatured on ice for 10 min, and incubated with suspensions of fibrocytes for 30 min at room temperature. After washing twice with DPBS, the fibrocytes were evaluated by flow cytometry.
3
Protein Lysate Immunoblotting Analysis
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Protein lysates were developed and exposed to sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS/PAGE), transported over polyvinylidene difluoride (PVDF) membranes, and blotted through standardized techniques, employing anti-collagen I antibody (1 : 100) (Millipore, USA) or anti-PI3K, anti-AKT, anti-phospho-AKT antibodies, anti-phospho-PI3K (1 : 100) (Abzoom Biolabs, USA). Anti-GAPDH antibodies (1 : 1000) (Abzoom Biolabs, USA) were used as a normalization control to ensure equivalent proteomic loading levels.
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