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
- - - - -

Mitochondrial swelling assay (PTP opening)


  • Please log in to reply
1 reply to this topic

#1 cardosopedro

cardosopedro

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 129 posts
3
Neutral

Posted 05 July 2013 - 09:52 AM

Hello.

Anyone here with experience with the mitochondrial swelling assay for the measurement of the opening of the permeability transition pore? I'm using the method that evaluates the decrease in absorbance at 540 nm. I'm working with a fungus and there's nothing in the literature about this. I get a "weird" profile (see attachment).

1) the "temporal dynamics" are quite different from what is observed in mammalian cells (normally I see 5 minute experiments and in my case I observed decrease in absorbance at least during the first half hour)

2) I get a very weird (but very reproducible) peak at 2-5mins after the addition of CaCl2. This is not inhibited by cyclosporin A, so I don't know what this is... any idea??

3) I get a decrease in absorbance after this, prolonged in time that is inhibited by cyclosporin A, so I guess this is the PTP opening.


Can someone please comment on this?


Thank you.
Posted Image

Attached Thumbnails

  • swelling.jpg

Edited by cardosopedro, 05 July 2013 - 09:52 AM.


#2 cardosopedro

cardosopedro

    Veteran

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 129 posts
3
Neutral

Posted 31 July 2013 - 04:33 AM

C'mon, help me! :)






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

©1999-2013 Protocol Online, All rights reserved.