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.
3 3 diaminobenzidine tetrahydrochloride dab
Manufactured by ZSGB-BIO
Sourced in China
3,3'-diaminobenzidine tetrahydrochloride (DAB) is a chromogenic substrate used in various immunohistochemical and enzyme-linked immunosorbent assay (ELISA) applications. It is commonly used as a peroxidase substrate to visualize the presence and localization of target proteins or antigens in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
Rat anti c3, by Abcam (2 mentions)
Goat anti rat secondary antibody, by ZSGB-BIO (2 mentions)
Bovine serum albumin (bsa), by neoFroxx (2 mentions)
Hematoxylin, by Solarbio (2 mentions)
Histostain plus kit, by ZSGB-BIO (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Biotinylated goat anti rabbit secondary antibody, by ZSGB-BIO (1 mentions)
Supersignal west femto maximum sensitivity substrate, by Thermo Fisher Scientific (1 mentions)
Upright fluorescence microscope, by Olympus (1 mentions)
Transparent dewaxing liquid, by Solarbio (1 mentions)
3 3 5 5 tetramethylbenzidine (tmb), by Tiangen Biotech (1 mentions)
Upright uorescence microscope, by Olympus (1 mentions)
Environmentally friendly transparent dewaxing liquid, by Solarbio (1 mentions)
7 protocols using 3 3 diaminobenzidine tetrahydrochloride dab
1
Neuroprotective Effects of SNX in Alzheimer's Disease
2
Immunohistochemical Detection of HEV, Tryptase, and 5-HT
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Tissue sections were prepared as previously described in this section, were deparaffinating, hydrated, water-bath heated for antigen retrieval and blocked with the addition of 3% hydrogen peroxide for immunohistochemistry. Afterward sections were incubated overnight with primary antibody monoclonal mouse anti-HEV ORF2, tryptase and 5-hydroxytryptamine antibody (1:200 dilution; Beijing Protein Institute, Beijing, China). Immunohistochemical staining was performed following the instructions that were included in the HistostainTM-Plus kit (ZSGB-BIO, Beijing, China). 3,3′-Diaminobenzidine tetrahydrochloride (DAB; ZSGB-BIO, Beijing, China) was applied for 5 min to visualize the antigen–antibody compound, Gill’s hematoxylin was applied as the background stain (Yang et al., 2018 (link)). The slides were observed under light microscope (Olympus Optical Co., Ltd., Beijing, China) and positive signals for HEV-ORF2 proteins were represented by a brown or yellow granular mass (Soomro et al., 2016 (link)). The positive staining intensity of ORF2, tryptase, and 5-hydroxytryptamine were measured as the ratio of the stained area to the total field assessed. Multiple views (three fields per section and five sections per animal) were randomly selected and analyzed.
3
Western Blot Immunodetection Reagents
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Horseradish peroxidase (HRP)-conjugated anti-rabbit and anti-mouse immunoglobulin G, and monoclonal antibody against β-Actin were obtained from Beyotime (Shanghai, China). BCA protein assay kit, SuperSignal West Femto maximum sensitivity substrate, and pierce protease and phosphatase inhibitor mini tablets, EDTA-free were obtained from Thermo Scientific (Waltham, MA). Rabbit polyclonal to LYRM4 was purchased from Abcam (Cambridge, UK). Streptavidin-biotin-peroxidase complex (SABC) and 3,3′-Diaminobenzidine tetrahydrochloride (DAB) were purchased from ZSGB-BIO (Beijing, China).
4
Immunohistochemical Analysis of Spinal Cord C3 Expression
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After antigen retrieval (see Immunofluorescence ), spinal cord sections were incubated in 3% H2O2. Next, sections were blocked with 3%BSA (BioFroxx) at room temperature for 1 h followed by incubation with rat anti-C3 (1:20, Abcam, UK) primary antibodies at 4 °C overnight. The next day, after washing with PBS three times, the tissues were incubated for 30 min at room temperature with goat anti-rat secondary antibody (1:200, ZSGB-Bio) and 3,3'-diaminobenzidine tetrahydrochloride (DAB, ZSGB-Bio). Next, sections were stained with hematoxylin (Solarbio) and differentiated with alcohol HCl. After rinsing with tap water, the slices were dehydrated in 70%, 80%, 90%, 95%, and 100% ethanol series, and made transparent with environmentally friendly transparent dewaxing liquid (Solarbio). Next, they were sealed with neutral gum and dried. Images were obtained using an upright fluorescence microscope (Olympus, Tokyo, Japan).
5
Immunohistochemical Detection of Hepatitis E Virus
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Livers, lungs, kidneys, spleens, lymph nodes, duodenums, jejunums, ileums, SR, and appendices were collected and fixed in 4% paraformaldehyde for 48 h. The fixed tissues were then processed routinely in paraffin and 4-mm sections were prepared. Monoclonal mouse anti-HEV ORF2 antibody (1∶300 dilution; Beijing Protein Institute, Beijing, China) was used as the primary antibody. The primary antibody was added to the sections and incubated at 37°C for 2 h. Immunohistochemical staining was performed according to the instructions of the Histostain™-Plus Kit (ZSGB-BIO, Beijing, China). 3,3′- Diaminobenzidine tetrahydrochloride (DAB; ZSGB-BIO, Beijing, China) was applied for 10 min to visualize the antigen–antibody reaction, and then Gill's hematoxylin was applied as the background stain. The primary antibody was replaced with phosphate-buffered saline in the negative control. The slides were then observed under an Olympus microscope (Japan).
6
Characterization of DC-SIGN and DC-SIGNR Antibodies
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Recombinant human DC-SIGN IgG-Fc fusion protein (rhDC-SIGN-Fc), DC-SIGNR IgG-Fc (rhDC-SIGNR-Fc) and anti-DC-SIGNR mouse monoclonal antibody (detection antibody) were purchased from R&D Systems (Minneapolis, MN, USA). Monoclonal anti-human DC-SIGN (capture antibody) produced in mouse was purchased from SIGMA-ALDRICH, INC, USA. A rabbit polyclonal antibody to DC-SIGN (detection antibody) and a monoclonal DC-SIGNR antibody (capture antibody) were purchased from Abcam, INC, Hong Kong, China. Another rabbit monoclonal antibody to DC-SIGNR was purchased from Epitomics, Hong Kong, China. Horseradish peroxidase (HRP) conjugated goat-anti-rabbit and anti-mouse antibodies, goat serum blocking reagent and 3,3′-diaminobenzidine tetrahydrochloride (DAB) were obtained from ZSGB-BIO (Beijing, China), and 3,3′,5,5′-tetramethylbenzidine (TMB) was purchased from TIANGEN BIOTECH CO, LTD, Beijing, China.
7
Immunohistochemical Analysis of C3 in Spinal Cord
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After antigen retrieval (see in Immuno uorescence), spinal cord tissues were incubated in 3% H 2 O 2 . Then, sections were blocked with 3%BSA (BioFroxx) at room temperature for 1 hour. The slices were incubated with rat anti-C3 (1:20, Abcam, UK) primary antibodies at 4℃ overnight. The next day, after washed with PBS 3 times, goat anti-rat secondary antibody (1:200, ZSGB-Bio) was used for incubating at room temperature for 30 minutes and stained with 3,3'-diaminobenzidine tetrahydrochloride (DAB, ZSGB-Bio). Next, sections were stained with hematoxylin (Solarbio) and differentiated with alcohol-HCl. After rinsing with tap water, slices were dehydrated in 70%, 80%, 90%, 95%, and 100% ethanol solution in turns, and made transparent with Environmentally friendly transparent dewaxing liquid (Solarbio). Then sealed with neutral gum and dried. Images were obtained with an upright uorescence microscope (Olympus, Tokyo, Japan).
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