H9c2 or HeLa cells were grown on coverslips. Cells were fixed with methanol for 15 min on ice and blocking/permeabilized with 5% goat serum (Sigma-Aldrich; Cat#G6767), 0.2% Triton-X100 (Sigma-Aldrich; Cat#X100) in PBS for 1 h at room temperature (RT) after 48-h transfection. Then, cells were incubated with primary antibody anti-RBM20 (rabbit, 1:400) in blocking buffer for 1 h at RT or overnight at 4°C. After washing, cells were incubated with Alexa flour-conjugated secondary antibody (1:1500; anti-rabbit, fluor 488, Cat#A32731; anti-rabbit, flour 568, Cat#A11036; Invitrogen) for 1 h at RT, washed with PBST, and mounted in SlowFade™ Gold Antifade Mountant with DAPI (Invitrogen; Cat#S36938). Cells were visualized and captured with a Zeiss LSM-710 confocal microscope with 405 and 488 nm laser lines, and Keyence BZ-X800 microscope (Osaka, Japan).
Lsm 710 confocal microscope
Manufactured by Keyence
Sourced in Japan
The LSM-710 is a confocal microscope designed for high-resolution imaging. It features a laser scanning system that allows for optical sectioning of samples, enabling the acquisition of three-dimensional data. The microscope is capable of capturing fluorescent and reflected light signals, making it suitable for a variety of applications in life science research and materials science.
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2 protocols using lsm 710 confocal microscope
1
Immunofluorescence Analysis of RBM20 in H9c2 and HeLa Cells
2
Immunofluorescence Assay for RBM20 in H9c2 and HeLa Cells
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H9c2 or HeLa cells were grown on coverslips. Cells were fixed with methanol for 15 min on ice and blocking/permeabilized with 5% goat serum (Sigma‐Aldrich; Cat#G6767), 0.2% Triton‐X100 (Sigma‐Aldrich; Cat#X100) in PBS for 1 h at room temperature (RT) after 48‐h transfection. Then, cells were incubated with primary antibody anti‐RBM20 (rabbit, 1:400) in blocking buffer for 1 h at RT or overnight at 4°C. After washing, cells were incubated with Alexa flour‐conjugated secondary antibody (1:1500; anti‐rabbit, fluor 488, Cat#A32731; anti‐rabbit, flour 568, Cat#A11036; Invitrogen) for 1 h at RT, washed with PBST, and mounted in SlowFade™ Gold Antifade Mountant with DAPI (Invitrogen; Cat#S36938). Cells were visualized and captured with a Zeiss LSM‐710 confocal microscope with 405 and 488 nm laser lines, and Keyence BZ‐X800 microscope (Osaka, Japan).
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