Immunofluorescence identification of Nestin protein expressed by NSCs
Take the 3rd generation neurosphere cultured NSCs, and fix for 4% paraformaldehyde for 20 min, break the membrane with 0.3% TritonX-100 for 15 min, block with 10% goat serum for 60 min at room temperature, add Nestin antibody (1:100), and incubate at 4℃ Overnight, add fluorescent secondary antibody, incubate at 37°C for 1h in the dark, add DAPI working solution dropwise, drop anti-fluorescence quenching solution 5 minutes later, mount the slide with neutral gum, and observe the Nestin expression of NSCs under a fluorescence microscope.