Jump to content

  • Log in with Facebook Log in with Twitter Log in with Windows Live Log In with Google      Sign In   
  • Create Account

Submit your paper to J Biol Methods today!
Photo
- - - - -

Hoechst33342/7-AAD staining

Hoescht 7-AAD Method FACS Dual Staining

  • Please log in to reply
No replies to this topic

#1 TinaTeaspoon

TinaTeaspoon

    member

  • Members
  • Pip
  • 3 posts
0
Neutral

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







Home - About - Terms of Service - Privacy - Contact Us

©1999-2013 Protocol Online, All rights reserved.