The FFPE tissues were deparaffinized and boiled in citrate buffer (MXB Cat#MVS-0100, Fuzhou, China) for antigen retrieval. Endogenous peroxidase was blocked by 3% H2O2. Then, slides were blocked in serum, incubated with anti-PLA2G2D antibody (diluted 1:50, Ancepta Cat# AP11151b) or anti-TMEM163 antibody (diluted 1:50, Invitrogen Cat# PA5-114329) at 37°C for 1h, incubated with an anti-rabbit secondary antibody, and visualized with diaminobenzidine (MXB Cat# DAB-2031, Fuzhou, China). The images of IHC staining were captured with a microscope.
Dab 2031
Manufactured by Beijing Strong Biotechnologies
Sourced in China
The DAB-2031 is a compact benchtop centrifuge designed for general laboratory applications. It features a robust construction, a brushless DC motor, and a digital display for precise speed and time control. The centrifuge can accommodate various sample tubes and microplates, making it a versatile tool for a range of laboratory tasks.
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Lab products found in correlation
5 protocols using dab 2031
1
Immunohistochemical Analysis of PLA2G2D and TMEM163
2
Immunohistochemical Analysis of FNDC5 in Lung
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Fresh tissue samples were collected from the inferior lobe of the left lung and fixed with 4% paraformaldehyde for 24 h. The samples were dehydrated using a series of ethanol, cleared using dimethyl benzene, and embedded in paraffin wax. Paraffin-embedded sections with a thickness of 4 µm were cut and stained with hematoxylin and eosin staining for 1 min at room temperature. The pathological changes in the lung tissues were observed using a light microscope (Zeiss Scope.A1; Zeiss AG; ×200 magnification). FNDC5 expression was detected using immunohistochemistry. Briefly, following deparaffinization and rehydration, antigen retrieval was performed by heat-mediated antigen retrieval with sodium citrate buffer (cat. no. DNS-0811, MXB Biotechnologies, pH 9.0) at 98°C for 15 min. Subsequently, the sections were stained with anti-FNDC5 (1:100; cat. no. BS-8486R, BIOSS) primary antibody at 4°C overnight. Subsequently, at room temperature, the sections were incubated with biotin-labeled goat anti-rabbit IgG (1:50; cat. no. SP KIT-C2, MXB Biotechnologies) for 30 min, followed by a reaction with diaminobenzidine (cat. no. DAB-2031, MXB Biotechnologies) for 3-5 min. Images were observed under an optical microscope (Zeiss Scope.A1; Zeiss AG; ×200 magnification).
3
Histological Analysis of Skin Wound Healing
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Animals were euthanized and skin samples were excised, the tissues were fixed with 4% paraformaldehyde for 24 hours, then dehydrated with 70%, 75%, 85%, 95%, 100% ethyl alcohol, covered with paraffin, before sectioning and histological analysis. Blocks were cut to expose wounded tissue near the center of each wound and then cut into 4 μm thickness and stained with hematoxylin and eosin.
Number of blood vessels containing red blood cells was counted over the entire wound area using three fields per section. The thickness of the new skin evaluated as follows: The middle part of the neonatal skin wound and the two ends near the edge of the wound were, respectively, selected to measure the thickness and then calculated the average.
The tissue sections were first deparaffinized and rehydrated prior to boil in a 100°C citrate buffer water bath for 30 minutes (R20902, YUANYE, China) and then quenched of endogenous peroxidize using blocker (KIT‐9720, MXB, China) for 10 minutes before incubating with CD31(28 364, abcam, UK) antibody overnight in 4°C, to allow visualization of the immunostaining, sections were incubated with the anti‐rabbit‐biotinylated secondary antibody for 45 minutes, and then incubated with Streptaridin‐Peroxidase for 20 minutes and DAB (DAB‐2031, MXB, China) and counterstained with hematoxylin. The proportion of CD31 positive signals is calculated by ImageJ software.
Number of blood vessels containing red blood cells was counted over the entire wound area using three fields per section. The thickness of the new skin evaluated as follows: The middle part of the neonatal skin wound and the two ends near the edge of the wound were, respectively, selected to measure the thickness and then calculated the average.
The tissue sections were first deparaffinized and rehydrated prior to boil in a 100°C citrate buffer water bath for 30 minutes (R20902, YUANYE, China) and then quenched of endogenous peroxidize using blocker (KIT‐9720, MXB, China) for 10 minutes before incubating with CD31(28 364, abcam, UK) antibody overnight in 4°C, to allow visualization of the immunostaining, sections were incubated with the anti‐rabbit‐biotinylated secondary antibody for 45 minutes, and then incubated with Streptaridin‐Peroxidase for 20 minutes and DAB (DAB‐2031, MXB, China) and counterstained with hematoxylin. The proportion of CD31 positive signals is calculated by ImageJ software.
4
Immunohistochemical Analysis of APP and BACE1
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Immunohistochemistry was performed on formalin-fixed, paraffin-embedded right brain hippocampal sections using three 5 μm coronal hippocampal sections per mouse. Primary antibodies against APP (1 : 300, MAB348, Millipore, USA) and BACE1 (1 : 700, ab183612, Abcam, USA) were diluted in PBS, and the sections were incubated at 4°C overnight, washed, and then stained with biotinylated secondary antibody (KIT-9706, KIT-9701, MXB, China) for 10 min. For detection of the positive expression, the sections were stained with DAB (DAB2031, MXB, China) for 10 min. Microscopy (BX53, Olympus, China) was performed, and images were obtained at 100x and 400x magnification, respectively.
5
MKK7 and c-Jun Expression in Hippocampus
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Immunohistochemistry was performed on formalin-fixed, paraffin-embedded right brain hippocampal sections using three 5-μm coronal hippocampal sections per mouse. Anti-mitogen-activated protein kinase kinase 7 (MKK7, 1:100) and c-Jun (60A8) rabbit mAb (1:70) were diluted in PBS, and the sections were incubated at 4°C overnight, washed, and then stained with biotinylated secondary antibody for 10 min. Positive expression was detected by staining the sections with DAB (DAB2031, MXB, China) for 10 min. Microscopy (BX53, Olympus, China) was performed, and images were obtained at 100 × and 400 × magnification. Information on the primary antibodies are displayed in Table 2 .
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