Hippuric acid was purchased from Sigma Aldrich (Cat. No. 112003). Antibodies were obtained for CD68 (Abcam; ab31630), phospho-SMAD2 (Cell Signaling Technologies; Cat. No. 3108), SMAD2 (Cell Signaling Technologies; Cat. No. 3103), transforming growth factor-β (TGF-β) (Cell Signaling Technologies; Cat. No. 3711), and collagen 1A (COL1A) (Cell Signaling Technologies; Cat. No. 91144). Harris Hematoxylin was purchased from Newcomer Supply (Cat. No. 1201). Xylene was purchased from Fisher Scientific (Cat. No. 05082-4). Eosin-Phloxine Working Solution was obtained from Newcomer Supply (Cat. No. 1082). Glacial acetic acid was purchased from Newcomer Supply (Cat. No. 10010 A). Sirius Red F3B (Cat. No. 36-554-8), a saturated aqueous solution of picric acid (Cat. No. P6744), and solid picric acid (Cat. No. 239801) were purchased from Sigma. Histoclear was obtained from National Diagnostics (Cat. No. HS-200). The DAB Staining kit (Ref: K4065), Protein Block reagent (Ref: X0909), and Antibody Diluent (Ref: 80809) were purchased from Dako. Cytoseal XYL Mounting Media was purchased from Thermo Scientific (Ref: 8312-4).
Cytoseal xyl mounting media
Manufactured by Thermo Fisher Scientific
Sourced in United States
Cytoseal XYL mounting media is a xylene-based solution used to permanently mount prepared microscope slides. It is designed to provide a clear, durable, and long-lasting seal for histological and cytological specimens.
Automatically generated - may contain errors
Lab products found in correlation
Bx40 microscope, by Olympus (4 mentions)
Eosin y, by Avantor (4 mentions)
Hematoxylin qs, by Vector Laboratories (4 mentions)
Positively charged slides, by Avantor (4 mentions)
Dp25 imaging system, by Olympus (3 mentions)
Catalyst one serum chemistry analyzer, by IDEXX (2 mentions)
Transwell permeable support, by Corning (2 mentions)
Transwell plate, by Corning (2 mentions)
Axio imager a2, by Zeiss (2 mentions)
Dab solution, by Vector Laboratories (2 mentions)
Xylene, by Thermo Fisher Scientific (1 mentions)
Antibody diluent, by Agilent Technologies (1 mentions)
Hippuric acid, by Merck Group (1 mentions)
Dab staining kit, by Agilent Technologies (1 mentions)
Smad2, by Cell Signaling Technology (1 mentions)
Phospho smad2, by Cell Signaling Technology (1 mentions)
Histoclear, by National Diagnostics (1 mentions)
Protein block reagent, by Agilent Technologies (1 mentions)
Bx43 microscope, by Olympus (1 mentions)
3 3 diaminobenzidine (dab), by Agilent Technologies (1 mentions)
14 protocols using cytoseal xyl mounting media
1
Immunohistochemistry of Fibrotic Markers
2
Histological Analysis of Liver Tissue
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Paraffin-embedded livers were cut into 4 μm sections and mounted onto positively charged slides (VWR, Radnor, PA, USA). Slides were deparaffinized and stained with Hematoxylin QS (Vector Laboratories, Burlingame, CA, USA) for 1 min followed by staining for 1 min with eosin Y (Amresco, Solon, OH, USA) and rinsed in 95% ethanol. The slides were then dipped into 100% ethanol and subsequently through two xylene washes. Coverslips were mounted onto the slides using CytoSeal XYL mounting media (ThermoFisher). The slides were viewed and imaged using an Olympus BX40 microscope with an Olympus DP25 imaging system (Olympus, Center Valley, PA, USA). Serum alanine aminotransferase (ALT) concentrations were measured using a commercially available kit from Sigma-Aldrich with all assays and subsequent analyses performed according to the instructions provided by the manufacturer.
3
Testicular Histological Analysis
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After dissection, testes were fixed in Bouin's fixative overnight at 4°C, dehydrated in a 30-90% ethanol gradient, cleared in xylene, and embedded in paraffin. Tissue blocks were sectioned at 7 µm, deparaffinized and rehydrated before staining. Briefly, slides were incubated with 1% periodic acid [Electron Microscopy Sciences (EMS), 19324-10] for 30 min at room temperature (RT), washed in running water for 5 min, then rinsed in deionized water. Slides were incubated with Schiff's reagent (EMS, 260582-05) for 30 min at RT and washed as described above before counterstaining with Hematoxylin Gill 2 for 30 s at RT. Slides were washed in running water for 1 min, dehydrated with ethanol, cleared with xylene, then mounted using Cytoseal XYL mounting media (Thermo Fisher Scientific, 22-050-262).
4
Histological Analysis of Paraffin-Embedded Liver Sections
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Paraffin-embedded livers were cut into 4-μm sections and mounted onto positively charged slides (VWR, Radnor, PA). Slides were deparaffinized and stained with Hematoxylin QS (Vector Laboratories, Burlingame, CA) followed by staining with eosin Y (Amresco, Solon, OH) and rinsed in 95% ethanol. The slides were then dipped into 100% ethanol and subsequently through two xylene washes. Coverslips were mounted onto the slides using CytoSeal XYL mounting media (ThermoFisher). The slides were viewed and imaged using an Olympus BX40 microscope with an Olympus DP25 imaging system (Olympus, Center Valley, PA). All images are taken as × 200 magnification.
Liver function was assessed by measuring serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels using a Catalyst One serum chemistry analyzer from IDEXX Laboratories, Inc. (Westbrook, MA).
Liver function was assessed by measuring serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels using a Catalyst One serum chemistry analyzer from IDEXX Laboratories, Inc. (Westbrook, MA).
5
Transwell Invasion Assay for 4T1 Breast Cancer Cells
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4T1 cells were plated at a density of 5 × 10 [5 (link)] cells/well in a Transwell plate (Corning, Corning, NY) and were exposed to 30 μM BP3 for 24 h. For chamber assays, 4T1 cells were grown to 70% confluence and pre-treated with either 0.1% DMSO or 30 μM BP3 for 2 weeks. The pretreated cells were trypsinized, centrifuged at 1000×g for 4 min, and brought to a concentration of 4.3 × 10 [5 (link)] cells/mL in serum-free media in Transwell permeable supports (Corning) above media containing 10% FBS. Following a 24-h incubation, Transwells were removed and fixed in 10% formalin for 10 min, stained with 10% Crystal Violet for 10 min, and rinsed with dH2O. Cells that did not migrate were removed from the upper surface by scrubbing with 1X PBS moistened cotton-tipped swab. The insert was removed from the chamber with a scalpel and mounted on a microscope slide with Cytoseal™XYL mounting media (Thermo Fisher Scientific, Waltham, MA). Images were captured with an Olympus BX41 light microscope using SPOT Software 5.1 (SPOT™ Imaging Solutions).
6
Histological Assessment of Liver Damage
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Liver damage was assessed by H&E staining according to previously published protocols.7 (link) Paraffin-embedded livers were cut into 4-μm sections and mounted onto positively charged slides (VWR, Radnor, PA). Slides were deparaffinized and stained with Hematoxylin QS (Vector Laboratories) for 1 minute followed by staining for 1 minute with eosin Y (Amresco, Solon, OH) and rinsed in 95% ethanol. The slides then were dipped into 100% ethanol and subsequently through 2 xylene washes. Coverslips were mounted onto the slides using CytoSeal XYL mounting media (Thermo Fisher Scientific, Waltham, MA). The slides were viewed and imaged using an Olympus BX40 microscope with an Olympus DP25 imaging system (Center Valley, PA).
Liver function was assessed by measuring plasma alanine aminotransferase and aspartate aminotransferase using the IDEXX Catalyst One machine from IDEXX Laboratories, Inc.
Liver function was assessed by measuring plasma alanine aminotransferase and aspartate aminotransferase using the IDEXX Catalyst One machine from IDEXX Laboratories, Inc.
7
Immunohistochemical Analysis of PAI-1 in Liver
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Formalin-fixed, paraffin-embedded liver sections were deparaffinized and re-hydrated through graded EtOH solutions. Sections were then incubated in 20% goat serum and 0.2% Triton-X100 for 1 h at room temperature followed by an overnight incubation with a 1:100 dilution of anti-PAI-1 antibody (MA5-17171, Thermo Fisher Scientific). Sections were thoroughly washed with Tris-buffered saline + 0.1% Tween-20, incubated with HRP-conjugated anti-mouse antibodies and then developed with DAB (Dako/Agilent, Santa Clara, CA). Sections were de-hydrated and mounted under Cytoseal XYL mounting media (Thermo Fisher Scientific). Photomicrographs were taken with an Olympus BX43 microscope equipped with CellSens version 2.3 software (Olympus Life Science, Waltham, MA).
8
Paraffin Embedding and Staining Protocol
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After dissection, testes were fixed in Bouin’s fixative overnight at 4°C, dehydrated in a 30–90% ethanol gradient, cleared in Xylenes, and embedded in paraffin. Tissue blocks were sectioned at 7 µm, deparaffinized, and rehydrated before staining. Briefly, slides were incubated with 1% periodic acid (Electron Microscopy Sciences (EMS); 19324–10) for 30 min at RT, washed in running water for 5 min, then rinsed in deionized water. Slides were incubated with Schiff’s reagent (EMS; 260582–05) for 30 min at RT and washed as described above before counterstaining with Hematoxylin Gill 2 for 30 s at RT. Slides were washed in running water for 1 min, dehydrated with ethanol, cleared with xylene, then mounted using Cytoseal XYL mounting media (Thermo Fisher Scientific; 22-050-262).
9
Transwell Invasion Assay for 4T1 Breast Cancer Cells
10
Nissl Staining for Tissue Sections
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Nissl staining was performed with the tissue sections mounted and dried overnight on the slides. The next day, the mounted sections were immersed in 100% xylene two times for five minutes each time. Subsequently, they were placed through a rehydration series of 3 immersions in 100% ethanol, followed by 95% ethanol, 70% ethanol, and tap water, with each step lasting 2 min. Afterward, the sections were immersed in cresyl violet solution for approximately two minutes. They were then dehydrated following a reversed order of the rehydration series and followed by a final immersion in xylene, with each step lasting up to one minute. Coverslips were mounted on the slides immediately after using Cytoseal XYL mounting media (Thermo Scientific, Waltham, MA, USA, #22-050-262).
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