48 hr after transfection, dissociated S2 cells were allowed to settle and adhere for 60 min on a glass coverslip. Cells were then washed twice with PBS, fixed in 4% formaldehyde (Electron Microscopy Sciences) in PBS, blocked and permeabilized by 1.5% normal goat serum (NGS) and 0.1% Triton X-100 in PBS, incubated with primary antibody in PBS containing 1.5% NGS and 0.1% Triton X-100 for 1 hr at room temperature, washed 3 times with 0.1% Triton X-100/PBS, incubated with secondary antibody (with or without Phalloidin) and washed with 0.1% Triton X-100/PBS. The stained cells were mounted using the Vectashield anti-fade mounting medium (H-1000) and imaged with a Zeiss spinning disc confocal microscope.
Anti fade mounting medium h 1000
Manufactured by Zeiss
Anti-fade mounting medium (H-1000) is a lab equipment product designed to preserve and protect fluorescent samples. It is formulated to reduce photobleaching and maintain the intensity of fluorescent signals during microscopic observation and imaging.
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2 protocols using anti fade mounting medium h 1000
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Immunofluorescence of Drosophila S2 cells
2
Immunofluorescence Staining of S2 Cells
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48 hours after transfection, dissociated S2 cells were allowed to settle and adhere for 60 min on a glass coverslip. Cells were then washed twice with PBS, fixed in 4% formaldehyde (Electron Microscopy Sciences) in PBS, blocked and permeabilized by 1.5% normal goat serum (NGS) & 0.1% Triton X-100 in PBS, incubated with primary antibody in PBS containing 1.5% NGS and 0.1% Triton X-100 for 1 hr at room temperature, washed 3 times with 0.1% Triton X-100/PBS, incubated with secondary antibody (with or without Phalloidin) and washed with 0.1% Triton X-100/PBS. The stained cells were mounted using the Vectashield anti-fade mounting medium (H-1000) and imaged with a Zeiss spinning disc confocal microscope.
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