Torularhodin (purity > 95%) was isolated and purified from the extract of Sporidiobolus pararoseus (JD-2 CCTCC M 2010326) according to a previously published method [10 (link)]. Animal diets were purchased from TROPHIC Animal Feed High-tech Co., Ltd. Commercial kits for serum concentration of total cholesterol (TC), low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL-Cc), and triglyceride (TG) were purchased from Jiancheng Technology Co. (Nanjing, China). Hematoxylin and eosin (H&E) were obtained from Servicebio Technology Co. (Wuhan, China). Antibodies of PPARα, PPAR-γ, CYP7A1, CPT1A, SLC27A4, and GAPDH were obtained from ABclonal Technology Co. Ltd. (Wuhan, China). Western fluorescence detection reagent was obtained from Beyotime Biotechnology (Shanghai, China).
Hematoxylin and eosin h e
Manufactured by Wuhan Servicebio Technology
Sourced in China
Hematoxylin and eosin (H&E) is a common staining technique used in histology and pathology laboratories. Hematoxylin stains cell nuclei blue, while eosin stains cytoplasmic structures and extracellular matrices pink. This staining method provides a basic visualization of tissue morphology and is widely used for routine histological analysis.
Automatically generated - may contain errors
Lab products found in correlation
Safranin o fast green, by Wuhan Servicebio Technology (2 mentions)
Cpt1a, by ABclonal (1 mentions)
Cyp7a1, by ABclonal (1 mentions)
Gapdh, by ABclonal (1 mentions)
Rm2235, by Leica (1 mentions)
Dmr 3000, by Leica (1 mentions)
Masson s trichrome staining, by Wuhan Servicebio Technology (1 mentions)
Periodic acid schiff, by Wuhan Servicebio Technology (1 mentions)
Trizol solution, by Takara Bio (1 mentions)
Goldner s trichrome, by Wuhan Servicebio Technology (1 mentions)
Toluidine blue, by Wuhan Servicebio Technology (1 mentions)
Citrate buffer, by Wuhan Servicebio Technology (1 mentions)
Alcian blue, by Wuhan Servicebio Technology (1 mentions)
Complete freund s adjuvant cfa, by Chondrex (1 mentions)
Caspase 3 activity assay kit, by Beyotime (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Solarbio (1 mentions)
Rpmi 1640 medium, by Solarbio (1 mentions)
Dna ladder, by Solarbio (1 mentions)
Agarose powder, by Biowest (1 mentions)
Sulfosuccinimidyl 4 n maleimidomethyl cyclohexane 1 carboxylate sulfo smcc, by Merck Group (1 mentions)
6 protocols using hematoxylin and eosin h e
1
Torularhodin Isolation and Purification from Sporidiobolus pararoseus
2
Histological and Immunohistochemical Analysis of Tissue Samples
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The acquired sample segments were submerged in 10% paraformaldehyde for 24 h. A paraffin microtome (RM2235, Leica, Wetazlar, GER) was used to slice samples embedded in paraffin blocks into 5-m thick sections.The slides were rehydrated dried and given a PBS washing. The slides were subsequently stained with hematoxylin and eosin (HE) (Servicebio, Wuhan, China) for histological analysis to gauge the percentage of inflammatory cells. Masson’s trichrome staining (Servicebio) was used to examine the density of collagen fibers and variations in collagen under a microscope.Subsequently, the slides were then treated with rabbit anti-ASPN antibody, rabbit anti-SMA antibody, and rabbit anti-collagen I antibody overnight at 4°C each and incubated with the secondary antibody at room temperature for 2 h. The slides were next stained with Meyer’s hematoxylin and 3,3-diaminobenzidine (DAB; Sigma, St. Louis, Missouri, United States) for 1–2 min (Sorabio, Beijing, P.R.C.). After blocking with neutral gum (Invitrogen, San Diego, CA, United States), the slides were examined under a microscope (Leica DMR 3000; Leica, Bensheim, Germany). ImageJ was used to quantitatively analyse the immunohistochemistry image data.
3
Detailed Characterization of TFRD Compound
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AFB1 (#2A1A08) was sourced from Pribolab Biological Engineering Co., Ltd. (Qingdao, China). TFRD (#K20798) had been acquired from Xi’an Kailai Biological Engineering Co., Ltd. (Xi’an, China). The compositional analysis of TFRD is shown in Figure S1 and Table S1 , and its main component was detected by liquid chromatography-mass spectrometry (LC-MS) analysis as Rutin (97.82%) [29 (link)]. Hematoxylin and eosin (HE) and Periodic Acid-Schiff (PAS) were purchased from Servicebio® (Wuhan, China). The 4% paraformaldehyde was purchased from Yantai Shuangshuang Chemical Co., Ltd. (Yantai, China). TRIzol solution was acquired from Takarabio Technology Co., Ltd. (Beijing, China; #AKF0726A).
4
Decalcification and Histological Analysis of Femoral Heads
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The femoral heads were decalcified with 14% EDTA solution (pH7.2) for 28 days. The decalcified samples were embedded in paraffin and cut into 4 μm sections. The Hematoxylin and Eosin (H&E) (Servicebio, Wuhan, China), Masson trichrome (MT) (Servicebio), Goldner’s trichrome (Servicebio), and Tartrate-resistant acid phosphatase (TRAP) (Servicebio) stainings were executed on the sections. The histological sections were observed with a light microscope (Olympus, Tokyo, Japan).
5
Histological Analysis of Decalcified Rat Knee
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The decalcified rat knee tissues were collected, dehydrated, and embedded in paraffin. They were cut into 4 μm-thick sections and stained with hematoxylin and eosin (HE) (Servicebio, China), safranin O-fast green (Servicebio, China), alcian blue (Servicebio, China), and toluidine blue (Servicebio, China). Additionally, immunohistochemical staining was performed using specific antibodies against ACAN, COL2A1, MMP3, and MMP13. The heart, liver, spleen, lung, and kidney of the rats were stained only with HE.
For the immunofluorescence staining, the sections were dewaxed, rehydrated, repaired with citrate buffer (Servicebio, China), blocked with 3% H2O2 for 30 min, and then blocked with 5% bovine serum albumin (Servicebio, China) for 1 h. Subsequently, the sections were incubated with the specific antibody against COL2A1 at 4°C overnight and incubated with the corresponding secondary antibody at room temperature in the dark for 30 min on the next day. The nuclei were stained with destination access point identifier dye (Servicebio, China), and the sections were observed under a fluorescence microscope.
For the immunofluorescence staining, the sections were dewaxed, rehydrated, repaired with citrate buffer (Servicebio, China), blocked with 3% H2O2 for 30 min, and then blocked with 5% bovine serum albumin (Servicebio, China) for 1 h. Subsequently, the sections were incubated with the specific antibody against COL2A1 at 4°C overnight and incubated with the corresponding secondary antibody at room temperature in the dark for 30 min on the next day. The nuclei were stained with destination access point identifier dye (Servicebio, China), and the sections were observed under a fluorescence microscope.
6
Synthesis and Characterization of Compounds for Rheumatoid Arthritis Study
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Oligonucleotides and peptide were synthesized by Sangon Biotech (Shanghai, China) and Huajin Biotechnology (Shanghai, China). Agarose powder was obtained from Biowest (Nuaillé, France). Sulfosuccinimidyl 4-[N-maleimidomethyl] cyclohexane-1-carboxylate (sulfo-SMCC) was supplied by Sigma (Darmstadt, Germany). EN was purchased from Shanghai CP Guojian Pharmaceutical (Shanghai, China). Cell counting kit 8 (CCK8), annexin V-FITC apoptosis staining/detection kit and caspase 3 activity assay kit were purchased from Beyotime Biotechnology (Shanghai, China). RPMI Medium 1640 was obtained from Beijing Solarbio Science & Technology (Beijing, China). Fetal bovine serum (FBS) was provided by BI (Kibbutz, Israel). Complete Freund's adjuvant (CFA) and immune grade chick type II Collagen were purchased from Chondrex (Woodinville, USA). Freund's adjuvant incomplete (IFA) was purchased from Sigma (Saint Louis, USA). Enzyme linked immunosorbent assay (ELISA) kits were purchased from MultiSciences Biotech (Hangzhou, China). DNA Ladder and 4′,6-diamidino-2-phenylindole (DAPI) were obtained from Beijing Solarbio Science & Technology (Beijing, China). Hematoxylin and eosin (H&E) and Safranin O-fast green were provided from Servicebio (Wuhan, China). All chemicals and materials were used as received without any further modifications.
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