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
- - - - -

metalloproteinase 2 handling


  • Please log in to reply
1 reply to this topic

#1 claire84

claire84

    Enthusiast

  • Active Members
  • PipPipPipPipPip
  • 34 posts
2
Neutral

Posted 24 February 2014 - 08:17 AM

Dear all,

I need your precious help!

I'm going to start some inhibition assays to test potential metalloproteinase 2 inhibitors. 

I bought  a batch of mmp2 with a concentration of 15 uM from "Enzo life sciences". For my inhibition assay I'll need an enzyme final concentration of 2 nM, so I decided to dilute the stock solution, to aliquot it and to store the aliquots at -80 °C. 

In the product data sheet they specified that procedures such as diluition of the enzyme followed by refreezing could lead to loss of activity. So I'm no more sure, that my ideas are correct and I don't know how to handle this enzyme.

 

Do you have some experience with this enzyme?

 

Please find attached the mmp2 product data sheet.

 

Every suggestions will be very appreciated!

 

thank you in advance

 

Chiara

 

 

 

 

 

 

Attached Files



#2 bob1

bob1

    Thelymitra pulchella

  • Global Moderators
  • PipPipPipPipPipPipPipPipPipPip
  • 5,663 posts
394
Excellent

Posted 24 February 2014 - 11:55 AM

In general freeze/thaw cycles can damage proteins, which would lead to loss of activity.  If the enzyme is stable at -20, you could add some glycerol to the storage buffer, as this will prevent the solution from freezing, and thereby preserve the activity.

 

Also, if you don't dilute, just aliquot and re-freeze you are less likely to see loss of activity (at least that is what is suggested by the data sheet).






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

©1999-2013 Protocol Online, All rights reserved.