Fluoromount g mounting solution

Manufactured by Southern Biotech

Sourced in United States

Fluoromount-G is a mounting solution designed for the preservation and preparation of fluorescent-labeled specimens for microscopy. It is a water-based, non-toxic medium that helps to maintain the fluorescence of labeled samples, enabling clear visualization and long-term storage.

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Lab products found in correlation

80i fluorescence microscope, by Nikon (2 mentions) Glass coverslip, by Avantor (2 mentions) Spinning disk confocal microscope, by Leica (2 mentions) Las af program software, by Leica (2 mentions) Confocal microscope lsm700, by Zeiss (1 mentions) E cadherin, by BD (1 mentions) Sp8 laser scanning confocal microscope, by Leica (1 mentions) Ab13970, by Abcam (1 mentions) Donkey serum, by Merck Group (1 mentions) Fluoview fv1000, by Olympus (1 mentions) Vinculin, by Merck Group (1 mentions) Pezgs0416, by Merck Group (1 mentions) Fluorescent dye conjugated secondary antibodies, by Jackson ImmunoResearch (1 mentions) Fluorescence microscope, by Zeiss (1 mentions) Fitc conjugated phalloidin, by Merck Group (1 mentions) β catenin, by BD (1 mentions) Anti ve cadherin, by BD (1 mentions)

8 protocols using fluoromount g mounting solution

Brain Tissue Preparation and Immunohistochemistry

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At 6 months after transplantation, animals were perfused transcardially with 4% paraformaldehyde. The brain was serially sectioned coronally and sagittally to 35 μm in thickness and subsequently stored in the preservation solution at −20°C.
Immunohistochemistry was performed as previously described (Liu et al., 2013b (link)). Sections were washed with PBS for 5 min three times, and permeabilized and blocked for 1 hr in 10% donkey serum and 0.2% Triton X-100 before being incubated in the primary antibody in 5% serum and 0.2% Triton X-100 at 4°C overnight. Sections were subsequently washed and stained with Alexa Fluor (Life) secondary antibodies and Hoechst in 5% donkey serum for 1 hr before being washed and mounted onto glass slides with Fluoromount-G mounting solution (SouthernBiotech). The primary antibodies used in this study are listed in Table S2. Images were visualized using a Nikon 80i fluorescence microscope (Nikon Instruments) and a Zeiss confocal microscope (LSM700, Zeiss instruments).

Huo H.Q., Qu Z.Y., Yuan F., Ma L., Yao L., Xu M., Hu Y., Ji J., Bhattacharyya A., Zhang S.C, & Liu Y. (2018). Modeling Down Syndrome with Patient iPSCs Reveals Cellular and Migration Deficits of GABAergic Neurons. Stem Cell Reports, 10(4), 1251-1266.

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Immunofluorescent Analysis of E14.5 Pancreata

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E14.5 pancreata were dissected in ice cold 1x PBS, then fixed in 4% paraformaldehyde (PFA) overnight at 4 C. After washing three times in 1x PBS, tissues were preserved in 30% sucrose in PBS at 4 C overnight and then embedded in Optimal Cutting Temperature (O.C.T.) compound (Tissue-Tek) and flash frozen prior to sectioning at 10 μm thickness.
For immunofluorescence staining, cryosections were washed 3 times in 1x PBS, permeabilized in 0.5% triton X-100 in PBS (PBT) for 10 min at room temperature (RT), and then blocked with 5% normal donkey serum (NDS) in 0.1% PBT for 1 h. Sections were stained overnight at 4 C using primary antibodies against GFP (1:500, Abcam Cat. ab13970), Chga (1:250, Abcam Cat. ab15160), or E-cadherin (1:100, BD Transduction Laboratories Cat. 610182). The next day, sections were washed three times in 1x PBS and then incubated with species-specific Alexa 488-, 555-, or 647-conjugated secondary antibodies and DAPI in 5% NDS in 0.1% PBT for 1 h at RT. Sections were washed three times in 1x PBS and covered in Fluoromount-G mounting solution (SouthernBiotech, Cat. 0100–01).
Images were captured with an SP8 Leica confocal laser scanning microscope. Maximum intensity Z-projections were then prepared using Image J software [36 (link)].

de la O S., Yao X., Chang S., Liu Z, & Sneddon J.B. (2023). Single-cell chromatin accessibility of developing murine pancreas identifies cell state-specific gene regulatory programs. Molecular Metabolism, 73, 101735.

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Immunofluorescence Imaging of Serine Metabolism

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Cells were cultured in conditional media containing 0.8 or 0 mM serine with an addition of 10% FBS on glass coverslips (VWR) in six-well dishes for 24 hr. Next, cells were fixed with 4% paraformaldehyde for 15 min at room temperature. After one wash in 1× PBS, cells were permeabilized in 0.15% Triton X-100 for 15 min at room temperature. Cells were then blocked in 10% BSA for 20 min at room temperature, followed by incubation in primary antibody solution (1:500 dilution in 10% BSA) for overnight at 4°C. After three washes using 1× PBS, cells were incubated in secondary antibody solution in 10% BSA and followed by another three times of wash. Cells were then mounted onto glass slides using Fluoromount-G mounting solution (Southern Biotech) for imaging acquisition on a spinning-disk confocal microscope (Leica) using a × 100/1.4NA oil or a ×40/1.25NA oil (Leica Plan Apochromat) objective. Images were acquired and processed using the Leica LAS AF program software.

Gao X., Lee K., Reid M.A., Sanderson S.M., Qiu C., Li S., Liu J, & Locasale J.W. (2018). Serine Availability Influences Mitochondrial Dynamics and Function through Lipid Metabolism. Cell reports, 22(13), 3507-3520.

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Immunofluorescence Staining of Organoids and Brain Slices

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The organoids and brain slices were washed three times with phosphate-buffered saline (PBS) and blocked for 1 h using 5% donkey serum (Millipore, Saint Louis, MO, USA) and 1% Triton X-100 (Biolink). The slices were incubated in primary antibody diluted with 0.2% Triton X-100 and 5% donkey serum at 4 °C overnight. The next day, the slices were washed three times with PBS and incubated in secondary antibody diluted in 5% donkey serum for 60 min at room temperature in the dark. Finally, the slices were washed and attached to glass slides with Fluoromount-G mounting solution (Southern Biotech). The primary and secondary antibodies are listed in Supplementary Table 2.

Dong X., Xu S.B., Chen X., Tao M., Tang X.Y., Fang K.H., Xu M., Pan Y., Chen Y., He S, & Liu Y. (2020). Human cerebral organoids establish subcortical projections in the mouse brain after transplantation. Molecular Psychiatry, 26(7), 2964-2976.

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Immunofluorescence Imaging of Serine Metabolism

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Quantifying fruM Expression in Drosophila Antennae

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Flies from the Or47b-CD2/CyO; fru^P1Gal4, 40xUAS-mCD8::GFP/TM6B stock were used for fru^M expression quantification. Fly heads were removed in PBT (phosphate buffered saline with Triton X) and fixed in 4% paraformaldehyde (PFA) for 1 hour, followed by three 15-min PBT washes. Antennae were removed from the head in PBT and fixed in 4% PFA for 0.5 hours, followed by three 15-min PBT washes. Antennae were incubated in primary antibody 1:200 mouse mouse-α-Rat-CD2 (Bio-Rad, MCA154GA) and 5% normal goat serum (NGS) at 4°C overnight, followed by three 15-min PBT washes, and then incubated in secondary antibody 1:1000 goat α-mouse-Cy3 and 5% NGS at 4°C overnight. Antennae were mounted using Fluoromount-G mounting solution (SouthernBiotech) and imaged. Confocal images were taken using the following confocal microscopes: Olympus FluoView FV1000 with laser power 488 = 17%, intensity = 710, pinhole = 105 μm, gain = 1, and offset = −1; Zeiss 510 with argon laser power = 35%, pinhole = 108 μm, detector gain = 800, amplifier gain = 1, and amplifier offset = −0.185.

Zhao S., Deanhardt B., Barlow G.T., Schleske P.G., Rossi A.M, & Volkan P.C. (2020). Chromatin-based reprogramming of a courtship regulator by concurrent pheromone perception and hormone signaling. Science Advances, 6(21), eaba6913.

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Immunofluorescence Staining of Neurons

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Neurons were fixed with 4% paraformaldehyde for 30 min. Neurons were washed with PBS for 5 min three times, permeabilized in 0.2% Triton X-100 for 10 min, and blocked for 1 hr in 10% donkey serum before being incubated in the primary antibody in 5% serum and 0.1% Triton X-100 at 4°C overnight. Cells were subsequently washed and stained with Alexa Fluor-conjugated secondary antibodies and Hoechst in 5% donkey serum for 1 hr before being washed and mounted onto glass slides with Fluoromount-G mounting solution (SouthernBiotech). The primary antibodies are listed in Table S2. Images were taken using a Nikon 80i fluorescence microscope (Nikon Instruments).

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Visualizing Cell-Cell Junction Proteins

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Cells (1 × 10⁴) were plated on fibronectin-coated 4-well chamber slides (PEZGS0416; Millipore, 5 µg/mL in serum-free DME). To determine localization of cell–cell junction molecules, 5 × 10⁴ cells were seeded and allowed to reach confluence. Cells were gently washed with PBS, fixed with cold 4% PFA for 10 min on ice, permeabilized with 0.1% Triton-X100 in PBS for 15 min at room temperature, and blocked with 1% BSA in TBS for 30 min. Slides were washed with TBS and incubated with primary antibodies (1:400) in 1% BSA in TBS at 4 °C overnight. Antibodies were used as follows: anti-VE-Cadherin (BD Bioscience), β-catenin (BD Biosciences), ZO-1 (Invitrogen), Vinculin (V4505, Sigma), and FITC-conjugated phalloidin (Sigma). The cells were rinsed with TBS and incubated with appropriate fluorescent-dye conjugated secondary antibodies (1:1000, Jackson ImmunoResearch) for 1 h at room temperature. The cells were then rinsed with TBS and incubated with DAPI (1:2000, D1306; Invitrogen) for 5 min and mounted on glass slides using a Fluoromount-G mounting solution (0100-01; SouthernBiotech, Birmingham, AL, USA). The cells were photographed with a Zeiss Fluorescence microscope (Axiophot, Zeiss, Germany) equipped with a digital camera.

Song Y.S., Jamali N., Sorenson C.M, & Sheibani N. (2023). Vitamin D Receptor Expression Limits the Angiogenic and Inflammatory Properties of Retinal Endothelial Cells. Cells, 12(2), 335.

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