We examined the cellular localization and expression levels of SOX2 and Nanog in parental SH-SY5Y and in multiple clones of MSDACs using Operetta (Perkin Elmer) high content and quantitative confocal imaging. Paraformaldehyde-fixed SH-SY5Y cells and MSDACs were permeabilized (0.25% Triton X-100), blocked (1% BSA in PBS), and labelled with rabbit polyclonal anti-SOX2 and anti-nanog antibody (1: 200, Santa Cruz). Then, they were tagged with Alexa Fluor 488 fluorochrome conjugated anti-mouse secondary antibodies (Abcam). The nucleus was counter-labeled with DAPI. After washing, the plates were analyzed in Operetta, at least eight fields/well and three wells/clone, with a minimum of 21 Z planes. Unstained controls were included for both cell lines. Columbus software (Perkin Elmer) was used for quantitative image analysis.
Alexa fluor 488 fluorochrome conjugated anti mouse secondary antibodies
Manufactured by Abcam
Alexa Fluor 488 fluorochrome conjugated anti-mouse secondary antibodies are fluorescent dye-labeled antibodies that specifically bind to mouse primary antibodies. They are used to detect and visualize target proteins or cells in various immunoassays and microscopy applications.
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2 protocols using alexa fluor 488 fluorochrome conjugated anti mouse secondary antibodies
1
Quantifying SOX2 and Nanog in Parental and MSDAC Cells
2
Quantitative Localization of RD3 Protein
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We examined the cellular localization and expression levels of RD3 in parental SH-SY5Y and MSDACs using Operetta (Perkin Elmer) high content and quantitative confocal imaging. Paraformaldehyde fixed SH-SY5Y and MSDACs were permeabilized (0.25% Triton X-100), blocked (1% BSA in PBS), and labelled with mouse monoclonal anti-RD3 (1: 200, Santa Cruz). Then, they were tagged with Alexa Fluor 488 fluorochrome conjugated anti-mouse secondary antibodies (Abcam). The nucleus was counter-labeled with DAPI. After washing, the plates were analyzed in Operetta with at least eight fields/well and three wells/clone, with a minimum of 21 Z planes. Unstained controls were included for both cell lines. Columbus software (Perkin Elmer) was used for quantitative image analysis.
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