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Immunocytochemistry

MS Manisha Singh
AK Anupama Kakkar
RS Rinkey Sharma
OK O. P. Kharbanda
NM Nitika Monga
MK Manish Kumar
SC Shantanu Chowdhary
BA Balram Airan
SM Sujata Mohanty
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The assay was performed as previously described20. Briefly, fixed cells were incubated overnight at 4 °C with primary monoclonal antibodies against Nestin (1:400), MAP2 (1:250) and TH (1:200) (Abcam, USA). After induction period, hMSCs were washed five times with PBS and incubated with fluoro isothiocyanate (FITC) conjugated secondary antibodies (1:500 dilution) for 1 hour at room temperature. Finally, after washing five tmes with PBS, cells were counter stained with 4′,6-diamidino-2-phenylindole (DAPI) to visualize the cell nuclei. Cells were washed thrice with PBS to remove excess DAPI stain. Stained cells were examined using a fluorescence microscope equipped with a digital camera (Nikon Eclipse 80i, Japan).

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