metalloproteinase 2 handling - (Feb/24/2014 )

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

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).