Clarion mounting medium

Manufactured by Merck Group

Sourced in United States

Clarion mounting medium is a laboratory product used to mount and preserve specimens for microscopic analysis. It serves as a transparent medium to mount and seal samples on microscope slides, allowing for clear visualization and long-term preservation of the specimens.

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Mounting medium (64 citations)

6 protocols using clarion mounting medium

Acetone-Based Protein Extraction and Analysis

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Acetone was purchased from Merk (Darmstadt, Germany). Tetramethylrhodamine isothiocyanate (TRITC), tetraethylrhodamine (Rhodamine B), dibutyl phthalate, phosphate-buffered saline (PBS) tablets (without Mg²⁺ and Ca²⁺), trichloroacetic acid (TCA) (>99%), dithiothreitol (DTT) (>99%), ammonium bicarbonate (99%), iodoacetamide (IAA) (>99%) and Clarion mounting medium, were purchased from Sigma-Aldrich Chemie (Steinheim, Germany). [Methyl-³H] thymidine was purchased from Amersham Biosciences (Buckinghamshire, UK). Peptide PMFIVNTNVPR was obtained from Peptide 2.0 (Chantilly, VA). Trypsin was purchased from Roche Applies Science (Mannheim, Germany). Trypsin/Lys-C mix was obtained from Promega (Madison, WI).

Karlsson I., Samuelsson K., Simonsson C., Stenfeldt A.L., Nilsson U., Ilag L.L., Jonsson C, & Karlberg A.T. (2018). The Fate of a Hapten - From the Skin to Modification of Macrophage Migration Inhibitory Factor (MIF) in Lymph Nodes. Scientific Reports, 8, 2895.

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Histological Staining of Transgenic Porcine Skin

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Formalin-fixed paraffin-embedded (FFPE) blocks of skin from transgenic and non-transgenic pigs were sectioned in 4 μm slices deparaffinized in xylene and rehydrated. The skin samples were placed in 0.1% Mayer's Hematoxylin (Sigma-Aldrich) for 15 min and rinsed in copious amounts of tap water. The slides were then successively dipped in 0.5% Eosin for 2 min and in ddH₂O until the Eosin stopped streaking. The samples were dehydrated in increasing concentrations of ethanol and lastly in xylene before being mounted with Clarion Mounting Medium (Sigma-Aldrich).

Staunstrup N.H., Stenderup K., Mortensen S., Primo M.N., Rosada C., Steiniche T., Liu Y., Li R., Schmidt M., Purup S., Dagnæs-Hansen F., Schrøder L.D., Svensson L., Petersen T.K., Callesen H., Bolund L, & Mikkelsen J.G. (2017). Psoriasiform skin disease in transgenic pigs with high-copy ectopic expression of human integrins α2 and β1. Disease Models & Mechanisms, 10(7), 869-880.

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Hematoxylin and Eosin (H&E) Staining Protocol

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H&E staining was performed on cell aggregate frozen sections as indicated by the manufacturer (Richard-Allen Scientific, MI, USA), using hematoxylin (7211, Richard-Allen Scientific, MI, USA) and eosin staining (7111, Richard-Allen Scientific, MI, USA). The slides were then cleared 3 times in 100% ethanol. Slides were then washed 3 times using Histo-Clear (HS-200, National Diagnostics, NC, USA) for 5 min, after which the slides were mounted using Clarion mounting medium (CO487, Sigma-Aldrich, MO, USA). Slides were then cured overnight at 37 °C before imaging.

Valdoz J.C., Franks N.A., Cribbs C.G., Jacobs D.J., Dodson E.L., Knight C.J., Poulson P.D., Garfield S.R., Johnson B.C., Hemeyer B.M., Sudo M.T., Saunooke J.A., Kartchner B.C., Saxton A., Vallecillo-Zuniga M.L., Santos M., Chamberlain B., Christensen K.A., Nordin G.P., Narayanan A.S., Raghu G, & Van Ry P.M. (2022). Soluble ECM promotes organotypic formation in lung alveolar model. Biomaterials, 283, 121464.

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Immunohistochemical Staining Protocol

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Cell aggregate sections were permeabilized using 0.1% Triton X-100 for 10 min and then blocked using Seablock buffer for 1hr at room temperature. Primary antibodies (Table S4) were diluted in 10% Seablock in PBST and incubated overnight at 4 °C. After intensive washing, HRP-conjugated secondary antibodies were then diluted to 1:500 in 10% Seablock PBST and incubated for an hour at RT. The chromogen was developed using a DAB kit (34002, Thermo Scientific, MO, USA) for 15 min at RT. Counterstaining for the nucleus was done using hematoxylin staining and bluing reagent. The slides were then cleared 3 times in 100% ethanol. Slides were then washed 3 times using Histo-Clear (HS-200, National Diagnostics, NC, USA) for 5 min, after which the slides were mounted using Clarion mounting medium (CO487, Sigma Aldrich, MO, USA). Slides were then cured overnight at 37 °C before imaging.

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Masson's Trichrome Staining Protocol

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Cell aggregate sections were processed following the protocol of Trichrome Stain (Masson) as indicated by the manufacturer (HT15–1 KT, Sigma-Aldrich, MO, USA), after which the slides were allowed to air dry. Slides were then washed using Histo-Clear (HS-200, National Diagnostics, NC, USA) for 5 min before being mounted using Clarion mounting medium (CO487, Sigma-Aldrich, MO, USA). Slides were then cured overnight at 37 °C before imaging.

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Cell Migration Assay Protocol

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Cells were collected and re-suspended in DMEM without serum at a density of 1×10 5 cells/mL. Next, 100 µL of cell suspension was added to the upper side of a transwell chamber with a pore size of 8 µm (Corning, Corning, NY, USA). Next, 600 µL of DMEM with 10% FBS was added to the lower chamber. The cells were allowed to migrate from the upper to the lower chamber for 18 h at 37°C. The membranes of the chambers were fixed with 4% paraformaldehyde. The cells that remained on the upper side of the membrane were gently removed using a cotton swab and cells left in the transwell chamber were stained with H&E staining solutions using the standard protocol. Afterwards, the membrane was cut out from the transwell chamber and mounted on a glass slide using Clarion Mounting Medium (Sigma-Aldrich, Saint Louis, MO, USA). Migrated cells were counted and imaged from each slide on a microscope with a 10X objective using a bright field filter (Axio Observer Z1, Carl Zeiss, Germany).

Ma S., Fu A., Chiew G.G, & Luo K.Q. (2017). Hemodynamic shear stress stimulates migration and extravasation of tumor cells by elevating cellular oxidative level. Cancer letters, 388.

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