Hello,

Could you please recommend DAPI concentration for nuclei staining in mesenchymal stem cells during chondrogenic differentiation?

I have used 1 μg/ml DAPI for 6 min, then as usual PBS washing. but I got fuzzy colour and lots of unspecific binding (but still fuzzy).

If that matters - the brief protocol:

- PBS washing 3x

- permeabilization 0.2% Triton X-100/PBS, 4 min, PBS washing 3x

- blocking 0,2% BSA/PBS 1h, PBS washing 3x

- staining of collagen I, II and X (each with a pair of antibodies, with 3x washing in PBS in between steps).

Thank you!

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