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Hoechst33342/7-AAD staining

Hoescht 7-AAD Method FACS Dual Staining

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#1 TinaTeaspoon

TinaTeaspoon

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Posted 10 September 2015 - 04:31 AM

Hello,

 

I am trying to dual stain with Hoechst33342 and 7-AAD. Does anyone have any experience using these dyes either individually or together? 

 

I set up an experiment following the nature protocols paper (link below) and did not detect any staining (even though I collected my media over 3 days and stored in the fridge, so I definitely had 'dead' cells).

 

http://www.nature.co...t.2006.458.html

 

The only parameters I changed was the amount of dyes added due to the recommendations on the datasheets which came with the dyes I purchased. I used 5ug Hoescht and 0.25ug 7-AAD per test instead of the 1ug Hoechst and 1ug 7-AAD suggested.

 

I could stain Hoechst for 20-30 min and then 7-AAD for 10 min all on ice also. Does any one have any other protocols they would recommend trialing or any other advice?

 

Best Wishes

 

TinaTeaspoon







Also tagged with one or more of these keywords: Hoescht, 7-AAD, Method, FACS, Dual Staining

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