The regenerated patellar tendon tissues were washed with PBS, fixed in buffered formalin, embedded in paraffin and sectioned for histological examination. Immunohistochemistry was done as described previously [26 (link)]. Briefly, after deparaffination, the sections were rehydrated, quenched of endogenous peroxidase activity and subject to antigen retrieval. After blocking with 5% normal donkey and goat serum, the sections were incubated with specific antibodies against Tnmd (sc-98875, Santa Cruz Biotechnology), Collagen I (AF7001, Affbiotech), GDF6 (bs-11843R, Bioss), Egr1 (bs-1076R, Bioss), OPN (bs-0026R, Bioss), OCN (bs-0470R, Bioss), MMP13(bs-0575R, Bioss) and Scx (sc-87425, Santa Cruz Biotechnology) at dilution of 1:200 at 4 °C overnight. Goat anti-rabbit horseradish peroxidase (HRP)-conjugated secondary antibody and donkey anti-goat horseradish peroxidase (HRP)-conjugated secondary antibody (Santa Cruz Biotechnology; dilution 1:200) were then added for 30 min, respectively. Afterward, the sections were rinsed, counterstained in hematoxylin or methylgreen, dehydrated with graded ethanol and xylene, and mounted with p-xylene-bis-pyridinium bromide (DPX) permount (Sigma Aldrich, St Louis, MO, USA). All incubation times and conditions were strictly controlled. The sections were examined under light microscopy (DMRXA2, Leica, Germany).
Bs 0026r
Manufactured by Bioss Antibodies
Sourced in United States
The Bs-0026R is a piece of lab equipment manufactured by Bioss Antibodies. It is designed for laboratory use. The core function of this product is to perform specific tasks within a laboratory setting. However, a detailed description cannot be provided while maintaining an unbiased and factual approach without further information.
Automatically generated - may contain errors
Lab products found in correlation
Bs 10196r, by Bioss Antibodies (2 mentions)
P xylene bis pyridinium bromide dpx permount, by Merck Group (1 mentions)
Dmrxa2, by Leica (1 mentions)
Sc 98875, by Santa Cruz Biotechnology (1 mentions)
Donkey anti goat horseradish peroxidase hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions)
Bs 0470r, by Bioss Antibodies (1 mentions)
Sc 87425, by Santa Cruz Biotechnology (1 mentions)
Df6421, by Affinity Biosciences (1 mentions)
Dab kit, by Maixin Group (1 mentions)
As014, by ABclonal (1 mentions)
Goat serum, by Solarbio (1 mentions)
P0216, by Beyotime (1 mentions)
Geldoc xr gel imaging system, by Bio-Rad (1 mentions)
Ipvh00010, by Merck Group (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Sa00001 2, by Proteintech (1 mentions)
4 protocols using bs 0026r
1
Histological Examination of Regenerated Patellar Tendon
2
Immunohistochemical Analysis of FSGS Samples
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Renal tissue sections including six patients of FSGS and three kidney para‐cancerous samples were obtained from the Second Affiliated Hospital of Guangxi Medical University. Ethical approval for this study was obtained from The Human Research Ethics Committee of the Second Affiliated Hospital of Guangxi Medical University. All patients provided written and verbal informed consent. Then tissue sections were incubated overnight at 4 °C with rabbit monoclonal antibodies against ALB (16475‐1‐AP; dilution 1 : 100; ProteinTech, Rosemont, IL, USA), APOE (AF5178; dilution 1 : 150; Affinity, Cincinnati, OH, USA), CLU (DF6421; dilution 1 : 100; Affinity), CST3 (DF6457; dilution 1 : 120; Affinity), SERPINA1 (DF6182; dilution 1 : 100; Affinity) and SPP1 (BS‐0026R; dilution 1 : 150; BIOSS, Woburn, MA, USA). This was followed by incubation with secondary antibodies at room temperature for 20 min. Tissue sections were dyed with hematoxylin and then counterstained, dehydrated and mounted. IHC results were analyzed using imagej (NIH, Bethesda, MD, USA).
3
Immunohistochemical Analysis of SP1, α-SMA, and OPN
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The 4-μm-thick sections were deparaffinized in xylene and hydrated using an ethanol-deionized water series. The sections were incubated with 3% H
2O
2 for 10 min at room temperature to block endogenous peroxidase activity, blocked with 5% goat serum (Solarbio), and then incubated with anti-SP1 antibody (1:200, bs-0975R; Bioss), anti-α-SMA antibody (1:200, bs-10196R; Bioss) and anti-OPN antibody (1:200, bs-0026R; Bioss) at 4°C overnight and then with HRP-labeled secondary antibody (1:500, AS014; ABclonal, Wuhan, China) at room temperature for 1 h. The sections were stained with a DAB kit (Maixin, Fuzhou, China) according to the manufacturer’s instructions. The sections were then counterstained with hematoxylin for 5 min at room temperature to visualize the nuclei and observed under the ECLIPSE Ci microscope (magnification×400). Brownish-yellow granules indicate immunoreactive (positive) cells, and the levels of positive protein expression were quantified using ImageJ 2x software.
2O
2 for 10 min at room temperature to block endogenous peroxidase activity, blocked with 5% goat serum (Solarbio), and then incubated with anti-SP1 antibody (1:200, bs-0975R; Bioss), anti-α-SMA antibody (1:200, bs-10196R; Bioss) and anti-OPN antibody (1:200, bs-0026R; Bioss) at 4°C overnight and then with HRP-labeled secondary antibody (1:500, AS014; ABclonal, Wuhan, China) at room temperature for 1 h. The sections were stained with a DAB kit (Maixin, Fuzhou, China) according to the manufacturer’s instructions. The sections were then counterstained with hematoxylin for 5 min at room temperature to visualize the nuclei and observed under the ECLIPSE Ci microscope (magnification×400). Brownish-yellow granules indicate immunoreactive (positive) cells, and the levels of positive protein expression were quantified using ImageJ 2x software.
4
Western Blotting Analysis of Protein Expression
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The cells were lysed using RIPA lysis buffer (P0013B; Beyotime) for total protein extraction. The proteins were quantified using a BCA kit (P0012; Beyotime), separated by 10% SDS-PAGE (P0012A; Beyotime), and transferred onto 0.45-μm polyvinylidene fluoride (PVDF) membranes (IPVH00010; Millipore, Billerica, USA). The membranes were blocked with 5% nonfat milk (P0216; Beyotime) for 2 h at room temperature and then incubated with primary antibodies against α-SMA (1:1000, bs-10196R; Bioss, Beijing, China), SM22α (1:1000, 10493-1-AP; ProteinTech, Rosemont, USA), CNN1 (1:1000, bs-0095R; Bioss), OPN (1:1000, bs-0026R; Bioss), Collagen I (1:1000, 14695-1-AP; ProteinTech) and β-actin (1:2000, 20536-1-AP; ProteinTech) overnight at 4°C. The membranes were then incubated with goat anti-rabbit IgG (1:5000, SA00001-2; ProteinTech) at 37°C for 1 h. An ECL chemiluminescence kit (P0018S; Beyotime Institute of Biotechnology) was utilized for chemiluminescence detection with the Bio-Rad Gel Doc XR
+ Gel Imaging System (Bio-Rad, Hercules, USA). β-Actin served as an internal control. ImageJ 2x software was used to quantify the protein bands.
+ Gel Imaging System (Bio-Rad, Hercules, USA). β-Actin served as an internal control. ImageJ 2x software was used to quantify the protein bands.
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