HUVECs were plated in four-well glass chamber slides and cultured to confluence. After sequential treatment with the indicated stimulant, cells were washed in phosphate-buffered saline (PBS) three times, and were then fixed with 4% paraformaldehyde for 10 min. Cells were washed again with PBS, followed by blocking with 0.5% bovine serum albumin (BSA) at room temperature and incubation with the appropriate mouse monoclonal VE-cadherin antibody overnight at 4 °C. After three washes with PBS, cells were incubated with FITC-conjugated goat anti-mouse antibody at room temperature for 1 h in the dark. Cells were mounted using DAPI-containing mounting medium (Santa Cruz Biotechnology). Immunoreactivity signals were visualized by confocal laser scanning microscopy (LSM780; Carl Zeiss, Dresden, Germany).
Dapi containing mounting medium
Manufactured by Santa Cruz Biotechnology
Sourced in Japan, United States
DAPI-containing mounting medium is a fluorescent staining solution used in microscopy applications. It is designed to mount and preserve biological samples while staining the cell nuclei with DAPI, a fluorescent dye that binds to DNA. This product facilitates the visualization and analysis of cellular structures under a fluorescence microscope.
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Lab products found in correlation
3 protocols using dapi containing mounting medium
1
VE-Cadherin Immunofluorescence in HUVECs
2
Immunofluorescence Analysis of Pyroptosis Markers
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hASCs were plated in 4-well glass chamber slides (SPL Life Science, Pocheon-Si, Korea) and cultured. After treatment, the cells were washed three times using PBS and fixed in 4% paraformaldehyde for 10 min. The cells were washed again with PBS, followed by blocking with 0.5% bovine serum albumin (BSA) at room temperature and incubation with anti-caspase-1 (1:200, ab1872, Abcam), anti-caspase-4 (1:100, sc-56056, Santa Cruz Biotechnology), anti-GSDMD (1:100, sc-393581, Santa Cruz Biotechnology), anti-IL-1β (1:100, NB600-633, Novusbio) antibody overnight at 4 °C. After washing three times with PBS, the cells were incubated with FITC-conjugated goat anti-rabbit IgG and rhodamine-conjugated rabbit anti-mouse IgG at room temperature for 1 h in the dark. The cells were mounted using DAPI-containing mounting medium (Santa Cruz Biotechnology) and then signals were examined using an Olympus BX53TR microscope (Tokyo, Japan). All images were processed using the CellSens software (Olympus).
3
Immunofluorescence Assay for MYOT Expression
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The cells were cultured on microscopic glass slides and grown to a near-confluent state. Afterwards, the cells were fixed in 4% PFA in PBS for 10 min at room temperature, permeabilized in ice-cold acetone/methanol (1:1) for 10 min at -20°C, rinsed with PBS and blocked in 3% BSA for 45 min. Anti-MYOT primary antibody (mouse monoclonal anti-MYOT antibody, 1:200 Santa Cruz Biotechnology, Santa Cruz, USA) was used for detection along with the corresponding green dye-labelled secondary antibody (MFP488, donkey anti-goat IgG, 1:200, MoBiTec, Goettingen, Germany). Afterwards, the cells were washed three times with PBS and sealed with DAPI-containing mounting medium (Santa Cruz Biotechnology, Santa Cruz, CA, USA). The expression of MYOT was analysed under a fluorescence microscope (Zeiss Axio-Imager.Z1) by pseudo-colour representations of fluorescence intensity for DAPI at 365 nm excitation and 420 nm emission wavelengths (blue) and for MFP488 at 470 nm excitation and 525 nm emission wavelengths (green).
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