Paraffin sectioning was performed at 4 μM. After dewaxing in xylene and rehydration in gradient ethanol, hematoxylin-eosin staining (HE staining) was conducted with an HE staining kit (Beyotime, China, Cat# C0105). Periodic acid-Schiff staining (PAS) was performed with a periodic acid-Schiff staining kit (Solarbio, China, Cat# G1281). Masson’s trichrome staining was performed with a kit from Nanjing Jiancheng Bio., China (Cat# D026). The above staining procedure were done according to the manufacturer’s instructions. Tubular injury score was semiquantitatively calculated based on PAS staining according to the percentage of cortical tubular necrosis with an assigned value: 0, none; 1, 10%; 2, 10% to 25%; 3, 25% to 75%; and 4, >75% [16 (link)]. Immunohistochemical staining (IHC) was performed with Biotin-Streptavidin HRP-based SPlink Detection Kits (ZSGB-Bio, China, Cat# SP-9002) and all procedures followed the manufacturer’s instructions. Antibodies used in IHC were mouse anti-α-SMA (1: 100, Boster, China, Cat# BM0002) and mouse anti-fibronectin (1: 100, DHSB, USA, Cat# P1H11). Images were taken with a light microscope (Nikon Eclipse 80i, Japan) at 200× magnification.
Biotin streptavidin hrp based splink detection kits
Manufactured by ZSGB-BIO
Sourced in China
The Biotin-Streptavidin HRP-based SPlink Detection Kits are laboratory equipment used for the detection and quantification of target molecules in various biological assays. The kits utilize the high-affinity interaction between biotin and streptavidin, coupled with horseradish peroxidase (HRP) for signal amplification. This system provides a sensitive and reliable method for the detection of target analytes in a variety of experimental settings.
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Lab products found in correlation
Periodic acid schiff staining kit, by Solarbio (1 mentions)
Eclipse 80i, by Nikon (1 mentions)
He staining kit, by Beyotime (1 mentions)
Virtual slide microscope vs120 s6 w, by Olympus (1 mentions)
Fibronectin, by Abcam (1 mentions)
α sma, by Abcam (1 mentions)
Virtual slide microscope, by Olympus (1 mentions)
3 protocols using biotin streptavidin hrp based splink detection kits
1
Histological Assessment of Tubular Injury
2
Immunohistochemical Analysis of Inflammatory Markers in Kidney
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For immunohistochemistry, TNF-α, IL-1β, IL-6, and F4/80 were tested in kidney samples on slices. First, the slices were dewaxed in xylene and rehydrated in gradient alcohol before antigen repair (citrate buffer pH 6.0, 95 °C for 10 min). Next, immunohistochemical staining of the tissue was performed according to the kit instructions. Briefly, the slices were incubated at 4 °C overnight with corresponding primary antibody (TNF-α, Santa, sc-52,746, USA, 1:200; IL-1β, Santa, sc-12,742, USA, 1:200; IL-6, Santa, sc-532,296, USA, 1:200; F4/80, Santa, sc-377,009, USA, 1:200) in 1% bovine serum albumin (BSA). The next day, after washing three times with PBS, the slices were treated with Biotin-Streptavidin HRP-based SPlink Detection Kits (ZSGB-Bio, PV-6000, China) and stained the nucleus with hematoxylin. Ultimately, all the slices were photographed with Virtual Slide Microscope (VS120-S6-W, Olympus, Japan).
3
Immunohistochemical Analysis of Kidney Tissue
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Mouse kidney tissues were carefully isolated, fixed with 4% paraformaldehyde, and embedded in paraffin. Then, the paraffin-embedded specimens were sectioned, deparaffinized, and rehydrated with gradient ethanol and subjected to subsequent detections including HE, Masson trichrome staining, and immunohistochemistry. Immunohistochemistry was performed according to the following procedures: the sections were immersed in citric acid antigen repair buffer based on microwave-based antigen retrieval technology as described before [7 (link)]. Then, the sections were reacted with indicated primary antibodies including α-SMA (1 : 100, China, Boster), fibronectin (1 : 100, USA, Abcam) overnight at 4°C. Immediately after that, the sections were rinsed with PBS, incubating with Biotin-Streptavidin HRP-based SPlink Detection Kits (ZSGB-Bio, China), and color was developed with DAB. Finally, Counterstaining was performed with hematoxylin. Signals were detected with a Virtual Slide Microscope (VS120, Olympus, Japan).
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