Liver tissues were embedded in paraffin and 5‐µm‐thick sections were prepared. The sections were placed on slides and heated at 60°C for 2 hours. Following dewaxing, rehydration, the slides were incubated in boiled antigen retrieval buffer for 10 minutes. Then slides were blocked with goat serum for 15 minutes at room temperature. For double labelling, slides were incubated with desmin antibody (1:50; Proteintech, Rosemont, IL) plus proliferating cell nuclear antigen (PCNA) antibody (1:50; Santacruz Biotechnology, Santa Cruz, CA) at 4°C overnight, followed by incubation with fluorescein isothiocyanate (FITC)‐conjugated anti‐mouse IgG (1:200; Beyotime, Haimen, China) and Cy3‐conjugated anti‐rabbit IgG (1:200; Beyotime) for 90 minutes at room temperature. Finally, the slides were counterstained with 4′,6‐diamidino‐2‐phenylindole (DAPI) and observed under a fluorescence microscope (BX53; Olympus).
Fitc conjugated anti mouse igg
Manufactured by Beyotime
Sourced in China
FITC-conjugated anti-mouse IgG is a laboratory reagent used for the detection and quantification of mouse immunoglobulin G (IgG) in various immunoassays. It consists of FITC (fluorescein isothiocyanate) molecules covalently linked to anti-mouse IgG antibodies, which can bind to and label mouse IgG for visualization or analysis.
Automatically generated - may contain errors
Lab products found in correlation
Cy3 conjugated anti rabbit igg, by Beyotime (3 mentions)
Proliferating cell nuclear antigen antibody, by Santa Cruz Biotechnology (1 mentions)
Fluoview fv10i, by Olympus (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Abcam (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Blocking buffer, by Beyotime (1 mentions)
Sc 514, by Santa Cruz Biotechnology (1 mentions)
Nbp2 12446, by Novus Biologicals (1 mentions)
A0568, by Beyotime (1 mentions)
A0516, by Beyotime (1 mentions)
Anti collagen 1, by Abcam (1 mentions)
8 hydroxy 2 deoxyguanosine 8 ohdg, by Abcam (1 mentions)
Occludin, by Abcam (1 mentions)
Nf κb, by Abcam (1 mentions)
Anti β actin antibody, by Merck Group (1 mentions)
Hrp conjugated anti mouse igg, by Beyotime (1 mentions)
Il 1β, by Thermo Fisher Scientific (1 mentions)
Fluorescence microscopy, by Olympus (1 mentions)
6 protocols using fitc conjugated anti mouse igg
1
Liver Tissue Immunofluorescence Analysis
2
Immunohistochemical Analysis of AOPPs
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Deparaffinized slices were blocked in 5% bovine serum albumin in PBS for 1 h at room temperature and then incubated overnight at 4 °C with the mouse anti-AOPPs (1:200, Department of immunology, Southern Medical University) overnight. After washing and incubated with FITC-conjugated anti-mouse Ig-G (1:100, Beyotime, China) for 1 h, the slices were stained by DAPI (Abcam, Cambridge, UK). Reacted slides were mounted and cover slipped. Images were captured with an Olympus FluoView FV10i self-contained confocal laser scanning microscope system (Olympus America Inc., PA, USA).
3
Visualizing NLRP3 and Caspase-1 in LPS-Stimulated RAW264.7 Cells
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RAW264.7 (1×104 cells/ml) were stimulated with 1 µg/ml LPS for 4, 8, 12 and 24 h. Following treatment, the cells were incubated with fixing buffer and were blocked with blocking buffer (Shanghai Beyotime Institute of Biotechnology) for 30 min at room temperature. Subsequently, the cells were incubated with primary antibodies against rabbit anti-NLRP3 (1:1,000; NBP2-12446; Novus Biologicals, LLC) and mouse anti-caspase-1 (1:1,000; sc-514; Santa Cruz Biotechnology, Inc.) for 2 h at 4°C. The cells were incubated with secondary antibodies Cy3-conjugated anti-rabbit IgG (1:1,000; A0516; Shanghai Beyotime Institute of Biotechnology) and FITC-conjugated anti-mouse IgG (1:1,000; A0568; Shanghai Beyotime Institute of Biotechnology) for 1 h at room temperature following incubation with the primary antibody and washing. DAPI was used for staining nuclei for 10 min at room temperature. Finally, the stained cells were visualized under a fluorescence microscope (Olympus Corporation, Tokyo, Japan).
4
Rhubarb Granules Attenuate Collagen-Induced Damage
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Twenty-eight specified pathogen-free, 3-month-old, male Sprague Dawley (SD) rats, were used for this study. Each rat was housed in our animal facility under pathogen-free conditions and fed a standard laboratory diet with free access to water. The temperature was maintained at 18–22°C with a 12-h light/dark cycle. All animal procedures complied with government-published recommendations for the use of laboratory animals. The study was approved by the Institutional Ethics Review Boards of Guangdong Provincial Hospital of Chinese Medicine.
The anti-collagen-I, occludin, 8-hydroxy-2'-deoxyguanosine (8-OHdG), and NF-κB antibodies were procured from Abcam (Cambridge, UK), anti-β-actin antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA). FITC-conjugated anti-mouse IgG, cy3-conjugated anti-rabbit IgG, HRP-conjugated anti-mouse IgG, and rabbit IgG were purchased from Beyotime Biotechnology Company (Shanghai, China).
Rhubarb granules were purchased from Jiang Yin Tianjiang Pharmaceutical Company (Product Lot: 1303196) and were monitored for contaminants (heavy metals, pesticides, and mycotoxins) before formulation.
The anti-collagen-I, occludin, 8-hydroxy-2'-deoxyguanosine (8-OHdG), and NF-κB antibodies were procured from Abcam (Cambridge, UK), anti-β-actin antibody was obtained from Sigma-Aldrich (St. Louis, MO, USA). FITC-conjugated anti-mouse IgG, cy3-conjugated anti-rabbit IgG, HRP-conjugated anti-mouse IgG, and rabbit IgG were purchased from Beyotime Biotechnology Company (Shanghai, China).
Rhubarb granules were purchased from Jiang Yin Tianjiang Pharmaceutical Company (Product Lot: 1303196) and were monitored for contaminants (heavy metals, pesticides, and mycotoxins) before formulation.
5
Immunofluorescence Staining of MRP14 in Nthy-ori 3-1 Cells
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Cells were grown on poly-D-lysine-coated coverslips in RPMI 1640 medium containing 10% FBS. Before treatment, Nthy-ori 3-1 cells were starved overnight and then treated with IL-1β 20 ng/mL (Peprotech, Rocky Hill, NJ, USA) at the indicated time points. After stimulation, the cells were fixed for 15 min with 4% paraformaldehyde and permeabilised for 5 min with PBS containing 0.1% Triton X-100, and nonspecific binding sites were blocked by incubation with PBS containing 3% BSA. For immunofluorescence staining, cells were incubated with a mouse monoclonal antibody against MRP14 (1:50) (Santa Cruz), and FITC-conjugated anti-mouse IgG (1:400, Beyotime, Shanghai, China) was used as the secondary antibody. Images were then recorded by fluorescence microscopy (Olympus).
6
Immunofluorescence Analysis of Basal Cells
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After the first passage, BCs were seeded onto glass cover slips in a 24-well plate and incubated for 24 h. Cells were fixed in 4 % paraformaldehyde for 15 min and blocked with normal goat serum for 30 min at 37 °C. Cells were rinsed thoroughly with phosphate buffered saline (PBS) and incubated with primary antibody against Cytokeratin 5 (1:100; Abcam) and p63 (1:100; ZhongShan Golden Bridge Biotechnology) overnight at 4 °C. Slides were rinsed, incubated with TRITC conjugated anti-rabbit IgG (1:500; Beyotime, Beijing, China) and FITC conjugated anti-mouse IgG at room temperature for 30 min, counterstained with DAPI, and visualized with confocal fluorescence microscopy.
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