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!
Vatamanu Sergiu-Marian thank you, I ll try. 1 ug/ml always worked well with other cells, it is a problem just for MSCs.
1 ug/ml DAPI are always worked well for most cells. If it is not work, try hoechst 33342 in 1-2 ug/ml.
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