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
- - - - -

Ethidium bromide protocol


  • Please log in to reply
2 replies to this topic

#1 aks

aks

    member

  • Active Members
  • Pip
  • 8 posts
0
Neutral

Posted 15 June 2010 - 10:08 AM

Hi,
I am growing MCF-7 cell line. I have been trying to stain dead cells using ethidium bromide and I havnt been very successful. I found the protocol online but it doesnt seem to work. if there is any other method with which I can stain the cells please let me know. Thanks in advance.

Protocol
1. grow the cells on a cover slip coated with Ploy-L-Lysine
2. put one drop of ethidium bromide (5ug/ml) on the cells and incubate for 10 mins
3. rinse the cover slip with PBS
4. expose to UV light and the dead cells will be red in color.


and what wave length is ideal to excite Ethidium Bromide?

#2 doxorubicin

doxorubicin

    Veteran

  • Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 192 posts
15
Good

Posted 15 June 2010 - 07:19 PM

It sounds like you can just use trypan blue and a hemocytometer, as in normal cell counting.
If you want to use a flow cytometer, consider switching to propidium iodide, and this protocol (for either EtBr or PI) looks pretty good:

http://www.immun.pit...l... dyes I.doc

#3 zodiac1505

zodiac1505

    Enthusiast

  • Moderators
  • PipPipPipPipPip
  • 92 posts
3
Neutral

Posted 29 July 2010 - 05:53 AM

Hi,
I am growing MCF-7 cell line. I have been trying to stain dead cells using ethidium bromide and I havnt been very successful. I found the protocol online but it doesnt seem to work. if there is any other method with which I can stain the cells please let me know. Thanks in advance.

Protocol
1. grow the cells on a cover slip coated with Ploy-L-Lysine
2. put one drop of ethidium bromide (5ug/ml) on the cells and incubate for 10 mins
3. rinse the cover slip with PBS
4. expose to UV light and the dead cells will be red in color.


and what wave length is ideal to excite Ethidium Bromide?


If you want to look for dead cells by FACS, you could use annexin (early apoptosis marker) or PI (late apoptosis marker). If you need protocols on how to do it, just let me know :)




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

©1999-2013 Protocol Online, All rights reserved.