HSC-T6 was purchased from Beijing Friendship Hospital (Beijing, China). TGF-β1, LY-364947 [4-[3-(2-pyridinyl)-1H-pyrazol-4-yl]-quinoline HTS 466284 transforming growth factor β Type I receptor kinase inhibitor, molecular formula: C17H12N4, weight: 272.30, and purity: ≥95% (HPLC)], DMSO, and MTT were purchased from Sigma (St. Louis, USA). AST [molecular weight: 248.36386; purity: ≥98%] was purchased from Pure-One Bio Technology (Shanghai, China). Malotilate was purchased from Yabang Epson Pharmaceutical (Jiangsu, China). Kits of ALT, AST, TBil, Alb, MDA, GSH, SOD, COL-I, COL-III, α-SMA, MMP-2, TIMP-2, hydroxyproline, H&E staining, Masson's trichrome staining, Sirius red staining, and Annexin V-FITC/PI double staining were purchased from Jiancheng Bioengineering Institute (Nanjing, China). Antibodies against α-SMA, TGF-β1, TβR-I, Smad 2, p-Smad 2 (S465/467), Smad 3, p-Smad 3 (S423/425), Smad 7, and β-actin were purchased from Cell Signaling Technology (MA, USA). HRP-labeled secondary antibodies were purchased from ZSGB-BIO (Beijing, China). RevertAid™ First Strand cDNA Synthesis Kit and SYBR Green Real-Time PCR Kit were purchased from Thermo Fisher (MA, USA).
H e staining
Manufactured by Nanjing Jiancheng
Sourced in China
H&E staining is a commonly used histological staining technique that employs hematoxylin and eosin dyes to stain nucleic acids (blue/purple) and proteins (pink/red) in tissue samples. This staining method provides contrast between different cellular and tissue components, enabling visualization and identification of various cell types and structures under a microscope.
Automatically generated - may contain errors
Lab products found in correlation
Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (1 mentions)
Ly364947, by Merck Group (1 mentions)
Hrp labeled secondary antibody, by ZSGB-BIO (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Methyl thiazolyl tetrazolium (mtt), by Merck Group (1 mentions)
Tgf β1, by Merck Group (1 mentions)
Sybr green real time pcr kit, by Thermo Fisher Scientific (1 mentions)
Glutathione (gsh), by Nanjing Jiancheng (1 mentions)
Hydroxyproline, by Nanjing Jiancheng (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
Tgf β1, by Cell Signaling Technology (1 mentions)
Masson s trichrome staining, by Nanjing Jiancheng (1 mentions)
Annexin 5 fitc pi double staining, by Nanjing Jiancheng (1 mentions)
α sma, by Cell Signaling Technology (1 mentions)
Smad2, by Cell Signaling Technology (1 mentions)
Paraformaldehyde (pfa), by Sangon (1 mentions)
Masson trichrome staining, by Nanjing Jiancheng (1 mentions)
Paraformaldehyde (pfa), by Wuhan Servicebio Technology (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
3 3 diaminobenzidine (dab), by Vector Laboratories (1 mentions)
7 protocols using h e staining
1
Hepatic Stellate Cell Activation Assay
2
Histopathological Analysis of Rat Myocardial Injury
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The newly taken out rat heart tissues were put into 4% paraformaldehyde and fixed for 24 hours. The fixed specimens were taken out, dehydrated by gradient ethanol, transparent xylene, and embedded in paraffin. After the tissue was cut into thin slices, H&E staining (Jian Cheng, Nanjing, China) was performed, and the histopathological changes were observed under a light microscope. Myocardial histopathological injury score was used to score myocardial edema, interstitial inflammation, and bleeding 24 h after CLP. 0 score for no lesion, 1 score for lesion scope less than 25%, 2 scores for lesion scope 25%-50%, 3 scores for lesion scope 50%-75, and 4 scores for lesion scope greater than 75%. 10 high power fields were observed in each film, and the mean value was used as the pathological injury score.
3
Assessing Lung Inflammation and Fibrosis
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On days 3, 7, 14, and 21 post-BLM treatment, the lungs from mice in each treatment group were collected. The right lungs were frozen until further use, while the left lung was collected after inflating with 1 ml of 4% paraformaldehyde (Sangon, Shanghai, China) under constant pressure and placed in 4% paraformaldehyde. The left lung tissues were then embedded in paraffin blocks and cut into 5 μm sections for hematoxylin and eosin (H&E) staining or Masson trichrome staining to observe the inflammatory cell infiltration and collagen deposition respectively (NanJing Jiancheng Bioengineering Institute, Nanjing, China).
4
Lung Injury Morphology Analysis
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The mice were anesthetized and the left lobes of lungs were collected, fixed in 10% buffered formalin and embedded in paraffin. Lung tissue sections (5 μm thick) were performed. To observe the morphology of lung injury, H&E staining was used following the manufacturer’s protocols (Nanjing Jiancheng Bioengineering Institute, Nanjing, China). Quantification of the alveoli numbers and mean linear intercept (MLI) were calculated as previously reported.13 (link)
5
H&E Staining of Tumor Tissues
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Tissue morphology assessment was performed using H&E staining. Briefly, the tumor tissues were fixed in 4% paraformaldehyde, embedded in paraffin, and sectioned into 5-μm slices. Subsequently, the sections underwent H&E staining following the manufacturer’s protocol (Nanjing Jiancheng, Nanjing, Jiangsu, China).
6
Liver Histology Analysis in Mice
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Liver tissues from the mice were fixed in 4% paraformaldehyde for 12 h at room temperature and then rinsed for 24 h. Subsequently, the fixed liver tissues were dehydrated through a series of ethanol concentrations, including 75%, 80%, 95%, and two rounds of 100% ethanol, with each step lasting 30 min at room temperature. For clearing, the dehydrated tissues were immersed in xylene twice, each time for 30 min at room temperature. Afterward, the liver tissues were embedded in paraffin and sectioned to a thickness of 5 µm. These sections were subjected to hematoxylin and eosin (H&E) staining (Nanjing Jiancheng Bioengineering Institute, Nanjing, China) for histologic examination and subsequently observed and photographed under an optical microscope to assess liver injury.
7
Cardiomyocyte Morphology Analysis in Mice
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Mice heart tissues were flushed with ice-cold PBS thoroughly after sacrifice and divided into several parts. Left ventricules were fixed with 4% paraformaldehyde (ServiceBio) for 48 h, embedded in paraffin, and cut into serial 4 μm sections. HE staining (Jiancheng) and WGA staining (Sigma–Aldrich), which combined with immunofluorescence staining of cardiac troponin T (15513-1-AP; Proteintech Group) and 4′,6-diamidino-2-phenylindole (Beyotime), were used to analyze the morphology of cardiomyocytes and quantify transverse cardiomyocyte size. For immunohistochemical staining of mice heart tissues, we used primary antibodies against phosphorylated YB-1 (CSB-PA204680; CUSABIO), YB-1 (ab76149; Abcam), and phosphorylated ERK (No. 4376; Cell Signaling Technology). The isotype antibody was used as negative control. Corresponding horseradish peroxidase–conjugated secondary antibodies (ServiceBio) and DAB (Vector Laboratories) were used for detection. Images were captured by microscopes (Olympus) and analyzed by Image-Pro Plus software (X-ray Scan).
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