HEK293T cells are transfected in 6-well format via Polyfect transfection reagent (Qiagen) using the manufacturer's suggested protocol with 300 ng of each AsCas12a plasmid (or SpCas9–2xNLS) and 150 ng of crRNA (or sgRNA) expression plasmid on a cover slip. Forty eight hours following transfection, transfection media was removed, cells were washed with 1x PBS and fixed with 4% formaldehyde in 1xPBS for 15 min at room temperature. Following blocking (blocking solution: 2% BSA, 0.3% Triton X-100 within 1× PBS), samples were stained with mouse anti-hemagglutinin (Sigma, H9658, 1:500), and Alexa 488 donkey anti-mouse IgG (H+L; Invitrogen, A-21202, 1:2000), sequentially. VECTASHIELD Mounting Medium with DAPI (Vector Laboratories, H-1200) was used to stain the nuclei and to mount the samples on slide. Images were taken with Zeiss AxioPlan 2 IE Motorized Microscope System.
Axioplan 2 ie motorized microscope system
Manufactured by Zeiss
The AxioPlan 2 IE Motorized Microscope System is a research-grade microscope designed for advanced imaging applications. It features motorized components for automated operation and enhanced precision. The system provides high-quality optics and illumination for a wide range of microscopy techniques.
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Lab products found in correlation
2 protocols using axioplan 2 ie motorized microscope system
1
HEK293T Cell Transfection and Imaging
2
Immunofluorescence Imaging of Cas9-Expressing Cells
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HEK293T cells are transfected in six-well format via Polyfect transfection reagent (Qiagen) using the manufacturer’s suggested protocol with 300 ng Cas9–Cas9 fusion expression plasmid and 150 ng of each sgRNA expression plasmid on a cover slip. 48 h following transfection, transfection media was removed, cells were washed with 1× PBS and fixed with 4% formaldehyde in 1× PBS for 15 min at room temperature. Following blocking (blocking solution: 2% BSA, 0.3% Triton X-100, within 1× PBS), samples were stained with mouse anti-hemagglutinin (Sigma, H9658, 1:500), and Alexa 488 donkey anti-mouse IgG (H+L; Invitrogen, A-21202, 1:2000), sequentially. VECTASHIELD mounting medium with DAPI (Vector Laboratories, H-1200) was used to stain the nuclei and to mount the samples on slide. Images were taken with Zeiss AxioPlan 2 IE Motorized Microscope System.
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