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
- - - - -

precipitation of pepstatin A


  • Please log in to reply
3 replies to this topic

#1 yobou

yobou

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 89 posts
1
Neutral

Posted 02 July 2009 - 08:12 AM

Dear all
when I add different concentrations of pepstatin A (10, 25, 50 microM final, stock was dissolved in DMSO) to DMEM it is precipitated and shaking the culture dish does not help. any recommendation?
thanks

#2 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,788 posts
132
Excellent

Posted 02 July 2009 - 10:04 AM

we always make up our pepstatin in methanol.

most of the literature from the suppliers recommend methanol or ethanol (sigma is the only company i found that mentions dmso).

the companies say that the working concentration range is 0.5-1 ug/ml (sigma says 1uM). you may be trying to put too much in to your medium.
talent does what it can
genius does what it must
i do what i get paid to do

#3 yobou

yobou

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 89 posts
1
Neutral

Posted 02 July 2009 - 12:28 PM

thanks mdfenko, but according to sigma data sheet they mentioned that (At 25 mg/mL DMSO Pepstatin A forms a clear, faint yellow solution). So do I have to purchase a new one?
As for the concentration used according to this paper: http://www.pubmedcen...bmedid=19425170
they used pepstatin A at 10 μg/ml final. and in another cancer research paper they used 100 microM concentration final. Do you use it to inhibit cell death or inhibit autophagy (in combination with E64D)?
thanks

#4 mdfenko

mdfenko

    an elder

  • Active Members
  • PipPipPipPipPipPipPipPipPipPip
  • 2,788 posts
132
Excellent

Posted 06 July 2009 - 07:50 AM

we were using pepstatin, at 1ug/ml [final], as part of an inhibitor panel for the proteases with which we worked.

i don't know if you have to buy more (did you solubilize the whole thing?). i would.

you could try to slowly increase the concentration of pepstatin in your medium.
talent does what it can
genius does what it must
i do what i get paid to do




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

©1999-2013 Protocol Online, All rights reserved.