Formalin-fixed tissue was embedded in paraffin (Thermo Fisher Scientific), sliced into 5-μm sections, dewaxed in serial xylene (Thermo Fisher Scientific), and rehydrated through graded ethanol solutions (Pharmco-Aaper, Shelbyville, KY). Antigen retrieval was performed by heating the sections in 0.01 M citrate buffer (pH 6.0) at 95°C for 10 minutes. Samples were incubated with primary antibodies against PCNA (Dako) or β-catenin (Cell Signaling Technology) overnight at 4 °C. Horseradish peroxidase (HRP)-conjugated secondary antibodies were then applied to the sections, followed by the chromogen 4-diaminobenzidine staining according to the instruction of HRP/DAB (ABC) Detection IHC kit (Abcam, Cambridge, MA). Sections were then counterstained with hematoxylin for 2 seconds and rinsed immediately by running water. Positive expressions of PCNA and βcatenin were observed under a light microscope (Olympus BX51) and quantified with Fiji software.
βcatenin
Manufactured by Olympus
Sourced in Japan
βcatenin is a key component of the Wnt signaling pathway, a fundamental cellular process involved in various aspects of development and homeostasis. It functions as a transcriptional co-activator, regulating the expression of target genes. Detailed information about its specific applications or intended uses is not available.
Automatically generated - may contain errors
Lab products found in correlation
Paraffin, by Thermo Fisher Scientific (1 mentions)
Proliferating cell nuclear antigen (pcna), by Agilent Technologies (1 mentions)
Serial xylene, by Thermo Fisher Scientific (1 mentions)
Hrp dab abc detection ihc kit, by Abcam (1 mentions)
Proliferating cell nuclear antigen (pcna), by Olympus (1 mentions)
β catenin, by Cell Signaling Technology (1 mentions)
Laser confocal microscopy, by Olympus (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
γh2ax, by Leica (1 mentions)
β catenin, by Beyotime (1 mentions)
2 protocols using βcatenin
1
Immunohistochemical Analysis of PCNA and β-catenin
2
Quantification of DNA Damage and Wnt Signaling
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After treatment, cells were fixed with 4% formaldehyde for 20 min at room temperature. Cells were permeabilized by Triton X-100 and subjected to goat serum blocking (Beyotime Biotechnology, Haimen, China). Cells were then incubated with primary antibodies against γ-H2AX (1:100 dilution) (Santa Cruz, CA, USA) and β-catenin (1:200 dilution) (Beyotime Biotechnology, Haimen, China) overnight at 4 °C. Cells were rinsed with PBS twice and then incubated in the dark with respective secondary antibodies for 60 min and DAPI for 5 min (Beyotime Biotechnology, Haimen, China). Using a fluorescence microscope (Leica, Weltzlar, Germany), the number of γ-H2AX foci per cell was quantified. Then, β-catenin fluorescence was measured by laser confocal microscopy (Olympus, Tokyo, Japan).
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