The brain tissue from WT and APP/PS1 mice was formalin-fixed and embedded in paraffin. For IHC, the paraffin-embedded sections were deparaffinized in xylene and rehydrated in a graded series of ethanol before staining. After antigen retrieval and blocking, the sections were then incubated with anti-NR4A1 antibody at 4 °C overnight. The second day, sections were washed in PBS and incubated with a biotinylated secondary goat anti-rabbit antibody (ZsBio) for 30 min at 37 °C, and then incubated with an avidin-biotin peroxidase complex (ZsBio) for 30 min at 37 °C. The sections were washed in PBS and incubated with 3,3′-diaminobenzidine (DAB, ZsBio) for 3 min. Hematoxylin was used to counterstain nuclei. A LEICA DM6000B automatic microscope (Leica, Germany) was used to collect images.
Avidin biotin peroxidase complex
Manufactured by ZSGB-BIO
Sourced in China
The Avidin-biotin peroxidase complex is a laboratory reagent used for various immunodetection techniques. It consists of the glycoprotein avidin and the enzyme peroxidase, which form a stable complex. The complex binds to biotin-labeled antibodies or other biomolecules, allowing for the detection and visualization of targeted analytes in biological samples.
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Lab products found in correlation
4 protocols using avidin biotin peroxidase complex
1
Immunohistochemical Analysis of NR4A1 in AD Mouse Model
2
Quantification of Alpha-SMA Expression
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Briefly, after antigen retrieval, tissue sections were incubated with a rabbit polyclonal anti-α-SMA-antibody (1:200 dilution; Proteintech, Chicago, IL, USA) overnight at 4°C, followed by incubation with a biotinylated secondary antibody and an avidin-biotin peroxidase complex (ZSGB-Bio, Beijing, China). Then, the immune reactions were developed by adding DAB chromogen substrate solution (ZSGB-Bio, Beijing, China) to the slides. Harris hematoxylin was used for counterstaining. Negative controls were run in parallel with all reactions. All specimens were scored by a board-certified pathologist. The immunohistochemical staining scoring for α-SMA was evaluated as “low” or “high” expression, regarding the rate of positive cells for each sample as described previously 15 (link).
3
Immunohistochemical Staining Protocol
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Briefly, tissue sections were incubated with a polyclonal antibody overnight at 4°C after antigen retrieval, followed by incubation with a biotinylated secondary antibody and an avidin-biotin peroxidase complex (ZSGB-Bio, Beijing, China). Then DAB chromogenic substrate solution (ZSGB-Bio, Beijing, China) was added to the slides to form an immunoreaction. Harris hematoxylin was used for counterstaining. IHC results were interpreted by two independent pathologists who were blinded to the clinicopathological information.
4
IHC Analysis of QSOX1 Protein Expression in Tissues
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Briefly, after antigen retrieval, tissue sections were incubated with a rabbit polyclonal anti-QSOX1 antibody (1:200 dilution; Proteintech, Chicago, IL, USA) overnight at 4°C, followed by incubation with a biotinylated secondary antibody and an avidin-biotin peroxidase complex (ZSGB-Bio, Beijing, China). Then, the immune reactions were developed by adding DAB chromogen- substrate solution (ZSGB-Bio, Beijing, China) to the slides. Harris hematoxylin was used for counterstaining. Negative controls were run in parallel with all reactions. All specimens were scored by a board-certified pathologist. The scoring was determined as follows: i) the proportion of tumor cells with IHC staining for QSOX1 protein expression (0: no staining, 1 (Low): <33%, 2 (Intermediate): 33 to 66%, 3 (High): >66%), and ii) the intensity of the stain (0: negative, 1: weak, 2: moderate, 3: strong staining intensity). An overall staining score of ≤3 indicated low expression and a score of >3 was considered high expression.
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