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.
Biotinylated goat anti rabbit secondary antibody
Manufactured by ZSGB-BIO
Sourced in China
Biotinylated goat anti-rabbit secondary antibody is a laboratory reagent used to detect the presence of rabbit primary antibodies in various immunoassays. It is a conjugate of goat-derived antibodies that specifically bind to rabbit antibodies and a biotin molecule, which can be utilized to amplify the signal from the target antibody.
Automatically generated - may contain errors
Lab products found in correlation
Avidin biotin peroxidase complex, by ZSGB-BIO (1 mentions)
3 3 diaminobenzidine dab, by ZSGB-BIO (1 mentions)
Anti p erk1 2, by Abcam (1 mentions)
Anti erk1 2, by Abcam (1 mentions)
Anti cdk5, by Abcam (1 mentions)
Anti adam10, by Abcam (1 mentions)
Anti gsk3β, by Abcam (1 mentions)
Anti total tau, by Abcam (1 mentions)
Anti bace1, by Abcam (1 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Proteintech (1 mentions)
Ab32575, by Abcam (1 mentions)
Ab281583, by Abcam (1 mentions)
3 3 diaminobenzidine dab, by Boster Bio (1 mentions)
3 3 diaminobenzidine tetrahydrochloride dab, by ZSGB-BIO (1 mentions)
Nis elements imaging software, by Nikon (1 mentions)
5 protocols using biotinylated goat anti rabbit secondary antibody
1
Immunohistochemical Analysis of NR4A1 in AD Mouse Model
2
Antibody Sources for Western Blot Analysis
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The anti-APP C-terminal (A8717) antibody was from Sigma (Sigma, St. Louis, USA) and anti-APP (6E10) antibody was from Covance (Princeton, NJ, USA). The anti-total tau, anti-tau-pS396, anti-tau-pT231, anti-ADAM10, anti-BACE1, anti-p-ERK1/2, anti-ERK1/2, anti-GSK3β, anti-GSK3β (S9) and anti-CDK5 antibodies were purchased from Abcam (Abcam, Cambridge, UK). The anti-tau-pS262 antibody was obtained from Santa Cruz (Santa Cruz Biotechnology, California, USA). The anti-NR4A1 and GAPDH antibodies were ordered from Proteintech (Proteintech, Wuhan, China). The horseradish peroxidase (HRP)-conjugated secondary antibodies were from Proteintech (Proteintech). The biotinylated secondary goat anti-rabbit antibody was purchased from ZsBio (Zhongshan Golden Bridge Biotechnology Co., Ltd, Beijing, China).
3
Quantifying Angiogenesis and Smooth Muscle Actin
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The tissue slices were deparaffinized and rehydrated (detailed procedure as previously described in H&E staining). In order to retrieve antigen, samples were first immersed in 0.1 M citrate buffer at 96 °C for 10 min and later incubated in 5% BSA for 2 h. The slices were incubated with antibody against CD31 (1:100, Cat# ab281583, Abcam) and α-SMA (1:200, Cat# ab32575, Abcam) at 4 °C overnight. After rinsing with PBS and incubated with biotinylated goat anti-rabbit secondary antibody (Cat# PV-6001, ZSGB-BIO, China) for 2 h, the tissue slices were colored with 3,3-diaminobenzidine (DAB) (Cat# AR1022, Boster, China), stained with hematoxylin, dehydrated with a gradient ethanol series, soaked with xylene, and then sealed with resin. Finally, five random locations of each tissue slice were selected to count the new capillaries with endothelial cells positive for CD31 via microscope (400 ×) or to analyze the average optical density values for α-SMA expression at the indicated time points using Image-Pro Plus 6.0 Software.
4
Neuroprotective Effects of SNX in Alzheimer's Disease
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SNX was obtained from Zhongxin Pharmaceuticals (Tianjin, China). d -gal and vitamin E (VE) were purchased from YiFang Technology Co., Ltd. (Tianjin, China). Donepezil (DON) was purchased from a drug store (Tianjin, China). Malondialdehyde (MDA), superoxide dismutase (SOD), glutathione (GSH), and glutathione peroxidase (GPx) assay kits were obtained from Nanjing Jiancheng Bioengineering Research Institute (Nanjing, China). Anti-cleaved caspase-3, anti-nuclear factor κB (NF-κB), and anti-glial fibrillary acidic protein (GFAP) antibodies were purchased from Boster Biological Engineering Co., Ltd. (Wuhan, China). Biotinylated goat anti-rabbit secondary antibody and 3,3′-diaminobenzidine tetrahydrochloride (DAB) were purchased from ZSGB-BIO (Beijing, China). The other reagents were commercially available and of analytical purity.
5
Immunohistochemical Analysis of PCNA
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Paraffin sections were blocked with Dual Endogenous Blocking Reagent and 20% normal rabbit serum (ZSGB-Bio, Beijing, China) and incubated with monoclonal rabbit anti-mouse PCNA antibodies (Proteintech, Beijing, China) overnight at 4°C, followed by incubation with a biotinylated goat anti-rabbit secondary antibody (ZSGB-Bio, Beijing, China). Quantification of immunoreactive cells was performed with NIS-Elements imaging software (Nikon, Tokyo, Japan).
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