Paraffin-embedded tissue sections (4 μm) were subjected to antigen retrieval, incubation with anti-CD31 (ab28364, abcam), anti-VEGF-A (BA0407, Boster), anti-HIF-1A (PB0245, Boster), anti-CD34 (BA0532, Boster), or anti-TSP1 (ab93653, abcam) antibodies, followed by incubation with a peroxidase-conjugated goat anti-rabbit IgG (KIT-9706, Maixin-Bio) secondary antibody. Samples were developed with DAB or AEC and counterstained with Mayor's hematoxylin (AR0005, Boster). Image Pro Plus 6.0 software was used for quantitative analysis.
Ar0005
Manufactured by Boster Bio
Sourced in China
The AR0005 is a laboratory instrument designed for routine analysis. It features an automated sample handling system, a spectroscopic detection module, and data processing software. The core function of the AR0005 is to perform quantitative measurements on various sample types.
Automatically generated - may contain errors
Lab products found in correlation
Ar1022, by Boster Bio (2 mentions)
Sv0002 12, by Boster Bio (2 mentions)
Ar1000, by Boster Bio (2 mentions)
Ab28364, by Abcam (1 mentions)
Ba0407, by Boster Bio (1 mentions)
Pb0245, by Boster Bio (1 mentions)
Ba0532, by Boster Bio (1 mentions)
Kit 9706, by Maixin Group (1 mentions)
Ab105060, by Abcam (1 mentions)
Ab5675, by Merck Group (1 mentions)
Image pro plus 6, by Media Cybernetics (1 mentions)
Piezo1 antibody, by Affinity Biosciences (1 mentions)
α sma antibody, by Cell Signaling Technology (1 mentions)
Inverted fluorescence microscope, by Nikon (1 mentions)
P0083, by Beyotime (1 mentions)
Ar0009, by Boster Bio (1 mentions)
Ab15580, by Abcam (1 mentions)
Sv0004, by Boster Bio (1 mentions)
Pannoramic desk, by 3DHISTECH (1 mentions)
Biotinylated secondary antibody, by Wuhan Servicebio Technology (1 mentions)
8 protocols using ar0005
1
Immunohistochemical Analysis of Angiogenic Factors
2
Immunohistochemical Analysis of GADD45B
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For IHC analysis, 4 μm sections of paraffin-embedded human tissue specimens were performed by a PV-9003 Kit (ZSGBBIO, China) according to the manufacturer’s instructions. In brief, after three rounds of TO-type biotablet transparent agent dewaxing, dehydration by gradient ethanol, and antigen retrieval in 0.01 M citrate solution at 95°C for 20 minutes, the samples were treated with 3% H2O2 at room temperature for 20 minutes to remove the effect of endogenous peroxidase, then blocked with normal goat serum for 20 minutes at room temperature. Tissue sections were incubated with the primary antibody against GADD45B (1:250 dilution, Abcam, ab105060, UK) overnight at 4°C. Incubation with secondary antibody labelled with horseradish peroxidase polymer was carried out for 30 minutes at room temperature. Finally, the staining processes were performed with DAB chromogenic solution (AR1022, BOSTER, China), and haematoxylin (AR0005, BOSTER, China) was used to counterstain the nuclei. Rabbit IgG was used as an isotype control. Images were captured with a microscope and assessed with Image-Pro Plus as mean density = integrated optical density/area of DAB staining.19 (link)
3
Immunohistochemistry of mGluR5 in Brain
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Brain sections were microwaved in sodium citrate buffer at 100 °C for 30 min for antigen retrieval, and then sections were quenched for endogenous peroxidases in 3% H2O2 in methanol for 10 min before blocking with 5% BSA for 15 min at RT. The sections were incubated with anti-mGluR5 antibody (1:200, AB5675, Merck, United States) at 4 °C overnight. Subsequently, the sections were incubated with 1:250-diluted HRP-conjugated goat anti-rabbit antibody (SV0002-12, Boster, China) at 37 °C for 1 h and visualized by incubation with 3,3-diaminobenzidine tetrahydrochloride (DAB, AR1000, Boster, China). The slices were counterstained with hematoxylin (AR0005, Boster, China) for 1 min, dehydrated, and routinely mounted (Chen et al., 2020 (link)). Images were captured with an OLYMPUS BX41 microscopy with DP Controller software and quantified with Image-Pro Plus 6.0 software (Media Cybernetics).
4
Immunohistochemical Analysis of Aquaporins
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Brain tissue was fixed in 10% buffered formalin solution and paraffin sections (5 μm in thickness) were prepared routinely. Sections were quenched for endogenous peroxidases in 3% H2O2 in methanol for 10 min before blocked with 5% BSA for 15 min at RT. The sections were incubated with anti-AQP1 (1:50), anti-AQP4 antibody (1:100), anti-AQP9 antibody (1:100), at 4°C overnight. Subsequently, the sections were incubated with HRP-conjugated goat anti-rabbit antibody (Boster, SV0002-12) at 37°C for 1 h and visualized by incubation with 3,3-diaminobenzidine tetrahydrochloride (DAB) (Boster, AR1000). The slices were counterstained with haematoxylin (Boster, AR 0005), dehydrated, and mounted in permount. For detection of PrPSc, the brain slices were exposed to the buffer containing 6M GdnCl at RT for 1 h prior to the routine IHC process.
5
Immunohistochemical Analysis of Heart Tissue
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For heart tissue staining, the obtained tissues (including the normal control (NC), MI-7d and MI-28d heart tissues) were first fixed by 4 % paraformaldehyde and then embedded in paraffin for preservation. After a series of treatments, the samples were blocked with 5% bovine albumin (BSA; MP Biomedicals, USA) for 2 h and washed 3 times with PBS, and then added primary antibodies at 4 °C overnight, containing Piezo1 antibody (1:200; Affinity, DF12083, China), integrin β1 polyclonal antibody (1:1000; Proteintech, 12594-1-AP, China) and α-SMA antibody (1:1000; Cell Signaling Technology, 19245, USA). Secondary antibody (1:100; Dako, P0448, China) was added for 60 min. The current 3,3′-diaminobenzidine tertrahydrochloride (DAB) (Boster, AR1024, China) was used for chromogenic development for 15 s. Cell nuclei were stained with hematoxylin dye (Boster, AR0005, China). The images were captured with an inverted fluorescence microscope (Nikon, Japan) and analyzed by ImageJ 6.0 (National Institutes of Health, USA).
6
Immunohistochemical Profiling of Murine Tumor Biomarkers
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The biomarkers in nude mice transplanted tumors were detected by IHC methods. Cut the wax block into 4-μm wax slices, and use an oven at 60°C for 1–2 h. The sections were removed for dewaxing, hydration, and then antigen repair with improved sodium citrate antigen repair solution (P0083, Beyotime, China). Incubated for 30 min with inactivated endogenous peroxidase 3% H2O2 (SV0004, BOSTER, China) to avoid light. Afterwards, they were blocked with ready-to-use normal goat serum (AR0009, BOSTER, China) for 30 min. After shaking off the blocking solution, Ki67 (1:300, ab15580, abcam, United Kingdom), HMOX1 (Rabbit, 1:200, 10701-1-AP, proteintech, China), LC3 (Rabbit, 1:500, 14600-1-AP, proteintech, China), CTSL (Rabbit, 1:500, proteintech, China) were added and placed in a refrigerator at 4°C overnight for incubation. Polymeric HRP-labeled anti-rabbit/mouse IgG (SV0004, BOSTER, China) was added after washing with PBS and incubated for 30 min at room temperature in the dark. The color was developed with DAB chromogen solution (C02-12, ORIGENE, America). After color development, counterstain with hematoxylin (AR0005, BOSTER, China). Afterwards, it was blue with alkaline PBS. Finally, dehydration was performed and the slides were mounted. Scanning observations were performed using a Pannoramic Desk (3DHISTECH, Hungary).
7
Immunohistochemical Analysis of Liver Fibrosis
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For IHC analysis, the paraffin-embedded liver sections (4 μm) were pretreated with dewaxed, rehydrated and heat-induced antigen retrieval. Then, the sections were immersed in hydrogen peroxide (3%) for 10 min to neutralize endogenous peroxidase activities and sequentially incubated with goat serum (5%) for 30 min to block non-specific binding sites of the tissues. Next, after immunostaining overnight at 4°C with antibodies against collagen I (1:500; Cat. no. GB11022-1), α-SMA (1:500; Cat. no. GB13044), the slides were immunostained with a biotinylated secondary antibody (1:100; Cat. no. G1216) for 1 h at room temperature. The primary antibodies and biotinylated secondary antibody used in IHC staining were purchased from Wuhan Servicebio Technology Co., Ltd (Wuhan, China). DAB chromogenic reagent (AR1022; Boster; Wuhan, China) was added to visualize positive staining and hematoxylin (AR0005; Boster; Wuhan, China) was used to mark the nucleus. For semi-quantitative analysis of positive cells, images (20× magnification) were acquired by microscopy and analysed using ImageJ V1.8.0 software (National Institutes of Health).
8
Immunohistochemical Analysis of Murine Knee Cartilage
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Knee joint sections of mice were deparaffinized and moved in 2 changes of xylene each for 10 min, hydrated in a series of ethanol (100% ethanol, 95% ethanol, followed by one shift of 70% ethanol), and rinsed twice in PBS. The antigen was retrieved by retrieval kit (AR0022; Boster) for the sections (30min; 37 °C). H2O2 (0.5%) was used to quench endogenous peroxidases for 10 min. The tissue sections were incubated with selected primary antibodies (overnight; 4 °C). Slides were rinsed in PBS containing 0.1% tween 20 (PBST, catalog P1397, Millipore Sigma) and antigen blocked with 5% normal goat serum. Next, overnight incubation at 4 °C with rabbit anti-mouse ACAN (1:200 dilution, catalog DF7561, Affinity), mouse anti-mouse Runx2 (1:200 dilution, Affinity; AF5186), rabbit anti-mouse MMP-13 (1:200 dilution, catalog 18165-1-AP, Proteintech), HIF-1α (1:200 dilution, Affinity; AF1009) and ATF4 (1:200 dilution, Affinity; DF6008). Slides were incubated with anti-rabbit secondary antibody for 30 min at room temperature, 3 times PBS rinsed for 5 min each, followed by two washes in deionized water for 5 min each. A 3-min incubation detected antibody binding to ACAN, MMP13, Runx2, HIF-1α, and ATF4 antigen with DAB peroxidase substrate (catalog zli90181, OriGene). Mayer's hematoxylin solution (catalog AR0005, BOSTER) was used to counterstain nuclei for 10–15 s.
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