I'm planning to stain fixed Vero nuclei with Hoechst 33342. The protocol manual provides quite a range for both concentration (0.2 - 2 µg/ml) and incubation time (1 - 15 min) when staining fixed animal cells. Has anyone found a working concentration and incubation time when staining fixed Vero cells with Hoechst 33342?
It's highly unlikely you will ever over-stain with Hoechst because there are limited binding sites for the dye.
Generally speaking, a concentration of 1-10 ug/mL and 15 minutes at RT in PBS or your detergent-free buffer of choice will work for virtually all fixed cells.
It's highly unlikely you will ever over-stain with Hoechst because there are limited binding sites for the dye.
Generally speaking, a concentration of 1-10 ug/mL and 15 minutes at RT in PBS or your detergent-free buffer of choice will work for virtually all fixed cells.
Thank you for your answer, Jordan! I'll go with 2 ug/mL and 15 minutes then.
edit: Looking good. :)
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