Fluoromount g

Manufactured by Zeiss

Sourced in Germany

Fluoromount G is a water-based mounting medium designed for fluorescence microscopy. It is formulated to preserve fluorescence signals and provide a clear, non-fading background. Fluoromount G can be used to mount a variety of fluorescently labeled specimens on microscope slides.

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Lab products found in correlation

4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions) Plan apochromat oil immersion objective, by Zeiss (1 mentions) Lsm 800, by Zeiss (1 mentions) Plan apochromat, by Zeiss (1 mentions) Ab9465, by Abcam (1 mentions) Ab56357, by Abcam (1 mentions) Ab11370, by Abcam (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Donkey anti goat alexa fluor 555, by Thermo Fisher Scientific (1 mentions) Axioplan 2 microscope, by Zeiss (1 mentions) Alexa fluor 488 goat anti rat, by Thermo Fisher Scientific (1 mentions) Axio imager, by Zeiss (1 mentions) Mab 6 11b 1, by Merck Group (1 mentions)

Close spelling variants of this product

Fluoromount (5 citations) DAPI-fluoromount G (5 citations)

5 protocols using fluoromount g

Immunofluorescence Assay for NF-κB p65 Localization

Cited 1 time

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The NF-κB p65 nuclear localization was detected by immunofluorescence assays using a fluorescence microscope. For this study, RAW 264.7 cells were cultured directly on glass coverslips in 24-well plates for 24 h. After stimulation with LPS in the presence or absence of EEPC, the cells were fixed with 4% paraformaldehyde in PBS, permeabilized with 0.2% triton X-100 in PBS, and blocked with 1.5% normal donkey serum. Polyclonal antibodies against anti-NF-κB p65 (1 μg/well) were applied for 1 h followed by an 1 h incubation with FITC-conjugated donkey anti-rabbit IgG. The position of the cell nucleus was determined with DAPI. After washing with PBS, the coverslips were mounted in Fluoromount-G, and the fluorescence was visualized using a fluorescence microscope (Carl Zeiss, Germany)
[33 (link)].

Jeong J.W., Lee H.H., Han M.H., Kim G.Y., Hong S.H., Park C, & Choi Y.H. (2014). Ethanol extract of Poria cocos reduces the production of inflammatory mediators by suppressing the NF-kappaB signaling pathway in lipopolysaccharide-stimulated RAW 264.7 macrophages. BMC Complementary and Alternative Medicine, 14, 101.

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Immunocytochemistry of Mesenchymal Stem Cells

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Mesenchymal stem cells were seeded on glass coverslips and incubated for 3 h at 37°C. Then, coverslips were washed with PBS and cells were fixed with 4% PFA and permeabilized with 0.1% Triton X-100. The blocking step was performed by incubating the cells with 5% FBS in PBS for 30 min at RT. Next, cells were washed and incubated with primary antibody and DAPI (1 μg/ml, Sigma) diluted in blocking buffer for 2 h. When required, incubation with secondary antibody, diluted in blocking buffer, was performed for 1 h. Coverslips were mounted in Fluoromount G and analyzed with the Spinning Disc (Zeiss) using 488, 543, and 633 nm excitation and a 20×/0.8 DIC objective lens. Images were processed with ZEN software.

Marini I., Siegemund M., Hutt M., Kontermann R.E, & Pfizenmaier K. (2017). Antitumor Activity of a Mesenchymal Stem Cell Line Stably Secreting a Tumor-Targeted TNF-Related Apoptosis-Inducing Ligand Fusion Protein. Frontiers in Immunology, 8, 536.

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Immunofluorescence Staining of Cardiomyocytes

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Cardiomyocytes were washed with DPBS and fixed with 4% paraformaldehyde for 15 min at room temperature. Samples were washed and permeabilized (0.5% Triton X-100 in phosphate buffered saline [PBS]), followed by incubation with blocking buffer (5% bovine serum albumin [BSA], 0.2% Tween-20 in PBS). Samples were incubated with primary antibody overnight at 4 °C. The following antibodies were used at the indicated dilution: anti-sarcomeric α-actinin (abcam, ab9465, 1:500); anti-cardiac troponin-I (abcam, ab56357, 1:500); anti-connexin 43 (abcam, ab11370, 1:250). Secondary antibody incubation was performed for 1 h at room temperature: donkey anti-mouse Alexa Fluor 647 (1:500); donkey anti-rabbit Alexa Fluor 488 (1:500); donkey anti-goat Alexa Fluor 594 (1:500). All antibodies were diluted in blocking buffer. Samples were counterstained with 4′,6-diamidino-2-phenylindole (DAPI) and mounted onto glass slides using Fluoromount G. Images were acquired using a laser scanning confocal microscope (LSM 800, Zeiss) equipped with a 20x Plan-APOCHROMAT and a 63x Plan-APOCHROMAT oil immersion objective (both Zeiss).

Esser T.U., Trossmann V.T., Lentz S., Engel F.B, & Scheibel T. (2021). Designing of spider silk proteins for human induced pluripotent stem cell-based cardiac tissue engineering. Materials Today Bio, 11, 100114.

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Immunohistochemistry of Sclerostin and CD31

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Tibiae were dissected and bones fixed in methanol‐free 4% PFA for 4 hours (4 °C). A 0.5 M EDTA solution was used for decalcification of the tibiae, followed by treatment with a solution of 20% sucrose, 2% polyvinylpyrrolidone in PBS at 4 °C. Tibiae were cryo‐embedded and 30 µm sections cut. Sections were permeabilized (0.3% Triton PBS) and blocked with 5% donkey serum, then incubated with sclerostin (SOST) primary antibody (R & D Systems, Minneapolis, MN, USA; 1:50) overnight (4 °C). Alexa Fluor 555 donkey anti‐goat (Invitrogen) was used as a secondary antibody (1:300). For CD31 primary (BD Biosciences, San Jose, CA, USA; 1:40) sections were incubated overnight (4 °C). Alexa Fluor 488 goat anti‐rat (Invitrogen) was used as secondary (1:400). Nuclei were stained with Hoechst 33342 (Invitrogen; 1:10,000) and sections mounted using Fluoromount G. Imaging was performed using a Zeiss Axioplan2 microscope.

Goring A., Sharma A., Javaheri B., Smith R.C., Kanczler J.M., Boyde A., Hesse E., Mahajan S., Olsen B.R., Pitsillides A.A., Schneider P., Oreffo R.O, & Clarkin C.E. (2019). Regulation of the Bone Vascular Network is Sexually Dimorphic. Journal of Bone and Mineral Research, 34(11), 2117-2132.

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Quantifying Ciliary Protein Levels in Cell Lines

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For immunofluorescence experiments in cell lines, cells were cultured on coverslips until confluent and starved for 48 h. To quantify ciliary GLI2 and GPR161 levels, cells were treated with 500 nM SAG or DMSO for 24 h after 24 h of serum starvation. Cells were fixed with 4% PFA for 10 min at room temperature. After blocking with 5% normal donkey serum, the cells were incubated with primary antibody solutions for 1 h at room temperature followed by treatment with secondary antibodies for 30 min along with DAPI. Primary antibodies used were against GPR161 (1:200, custom-made)^{21 (link)}, acetylated α-tubulin (mAb 6-11B-1, Sigma; 1:2000), GLI2 (1:500, gift from Jonathan Eggenschwiler)^{91 (link)}, pericentrin (611814, BD Biosciences; 1:500). Coverslips were mounted with Fluoromount-G and images were acquired with a Zeiss AxioImager.Z1 microscope using a 40x oil immersion objective lens.

Hoppe N., Harrison S., Hwang S.H., Chen Z., Karelina M., Deshpande I., Suomivuori C.M., Palicharla V.R., Berry S.P., Tschaikner P., Regele D., Covey D.F., Stefan E., Marks D.S., Reiter J., Dror R.O., Evers A.S., Mukhopadhyay S, & Manglik A. (2023). GPR161 structure uncovers the redundant role of sterol-regulated ciliary cAMP signaling in the Hedgehog pathway. bioRxiv.

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