Paraffin-embedded mouse lung tissue was sectioned at a thickness of 3 µm. Deparaffinized/rehydrated sections were autoclaved at 120 °C for 10 min in citrate buffer (pH 6.0) for antigen retrieval. Sections were blocked for endogenous peroxidase activity using 3% hydrogen peroxide in methanol for 10 min and exposed to a blocking solution (#03649-64, Nacalai Tesque) for 10 min before incubation overnight in a humidified chamber at 4 °C with primary antibodies or isotype control. Immune complexes were detected by incubation with Simple Stain MAX-PO (M) (#424131; Nichirei, Tokyo, Japan) or Simple Stain MAX-PO (R) (#414341, Nichirei) at room temperature for 1 h before applying chromogen detection using diaminobenzidine (#425011, DAB substrate kit, Nichirei). Counterstaining was performed with Mayer’s hematoxylin (#30002; Muto pure chemicals, Tokyo, Japan) before dehydration. Sections were clarified with xylene and mounted using Marinol (#4197193, Muto pure chemicals). Images were acquired with a BZ-X810 microscope (Keyence) and analyzed with Image J software.
Marinol
Manufactured by Muto Pure Chemicals
Sourced in Japan
Marinol is a laboratory equipment product designed for use in chemical analysis and research. It is a precision weighing device capable of measuring substances with high accuracy. The core function of Marinol is to provide reliable and consistent weight measurements for various samples and materials used in laboratory experiments and processes.
Automatically generated - may contain errors
Lab products found in correlation
Simple stain max po m, by Nichirei Biosciences (1 mentions)
Bz x810 microscope, by Keyence (1 mentions)
Mayer s hematoxylin, by Muto Pure Chemicals (1 mentions)
Sm 200r microtome, by Leica (1 mentions)
Ab8536, by Abcam (1 mentions)
Biotinylated secondary antibody, by Vector Laboratories (1 mentions)
Avidin peroxidase complex, by Vector Laboratories (1 mentions)
3 3 diaminobenzidine (dab), by Vector Laboratories (1 mentions)
Microm hm 650v, by Thermo Fisher Scientific (1 mentions)
Paraformaldehyde phosphate buffer solution, by Fujifilm (1 mentions)
Hematoxylin, by Sakura Finetek (1 mentions)
Ethanol, by Fujifilm (1 mentions)
5 protocols using marinol
1
Immunohistochemical Analysis of Mouse Lung Tissue
2
Histological Analysis of Mouse Eyes
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The eyes of the excised mice were fixed by TB-Fix [3.7% formaldehyde (Wako, 064-00406), 15% ethanol (Wako, 057-00456), 10% acetic acid (Wako, 017-00256)], as previously reported (Tokuda et al., 2018 (link)). Briefly, the eyes were kept overnight at 4°C in PBS containing 10% formaldehyde, 10% ethanol and 10% acetic acid. Fixed eyes were embedded with paraffin and sectioned to a thickness of 5 µm using a SM 200R microtome (Leica). Slices dried on glass slides were stained with H&E (Merck Millipore) after the paraffin was removed and dehydrated with alcohol and xylene. These sections were mounted using Marinol (Muto Pure Chemicals).
3
Immunohistochemical Analysis of CD34 Expression
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Sections of formalin-fixed, paraffin-embedded tissues were depleted of paraffin and hydrated. They were incubated with 3% H2O2 for 5 min, washed with 0.05% Tween in Tris-buffered saline (TBST), exposed to blocking buffer [5% goat serum and 0.5% bovine serum albumin in phosphate-buffered saline (PBS)] in PBS for 10 min at 24 degrees Celsius, and incubated with mouse anti-CD34 antibodies (ab8536; clone QBEND-10; Abcam, Cambridge, UK) for 60 min. They were then washed with TBST, incubated with biotinylated secondary antibodies (Vector Laboratories, CA, USA) for 10 min at room temperature, and washed again, after which immune complexes were detected using an avidin-peroxidase complex (Vector Laboratories) and 3,3'-diaminobenzidine (Vector Laboratories). The sections were counterstained with Mayer’s hematoxylin, dehydrated using a graded series of ethanol, and mounted in Marinol (Muto Pure Chemicals, Tokyo, Japan).
4
Golgi Staining of VCP Knock-In Mice
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For Golgi staining, VCPT262A-KI and C57BL/6 littermate mice were fixed by cardiovascular perfusion of fixation solution (4% paraformaldehyde and 20% glutaraldehyde in 0.05 M phosphate buffer). Brains were immersed in the same fixation solution for another 1 h at 4°C, washed with 0.1 M phosphate buffer, incubated with potassium dichromate solution (5% potassium dichromate, 4.875% chloral hydrate, 1.5% glutaraldehyde, and 2% PFA) for 7 d at room temperature, washed three times with ddH2O, and incubated in 0.7% silver nitrate solution for 7 d at room temperature. Then, the brain samples were dehydrated with ethanol (70% for 1 h, 90% for 1 h, 100% for 2 h, and 50% ether plus 50% ethanol for 1 h), and embedded in celloidin solution (0.25% for 1 h, 5% for 12 h, and 10% for 48 h) at room temperature. 50-μm sections prepared with a vibrating microtome (Microm HM650V; Thermo Fisher Scientific) were dehydrated with 70% ethanol (three times), anhydrous alcohol, and 3:1 anhydrous alcohol:chloroform (three times); cleared with xylene (three times), and coverslipped with Marinol (#2009-3; Muto Pure Chemicals).
5
Histological Analysis of Xenograft Tumors
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Skin samples from the xenografted tumor sites and other vital organs, including the lung, heart, kidney, liver, and alimentary tracts, were obtained from the mice, fixed with 4% paraformaldehyde phosphate buffer solution (#163–20,145, FUJIFILM Wako Pure Chemical Corporation [FUJIFILM]) for 24 hours at 4°C, and embedded in paraffin. The histopathological specimens were deparaffinized by immersion in xylene (#241–00091, FUJIFILM) for 10 minutes at room temperature, and rehydrated by immersion in ethanol (#057–00451, FUJIFILM). Hematoxylin (#6187‐4P, Sakura Finetek) and eosin (#8660, Sakura Finetek) were used for H&E staining following the manufacturer's protocols. The stained slides were dehydrated by immersion in ethanol followed by xylene. Glass coverslips with Marinol (#4197193; Muto Pure Chemicals) were used to cover the stained slides. H&E‐stained slides were examined using OLYMPUS cellSens Standard system (OLYMPUS).
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