Does glycerol intefere an enzymatic reaction ? - (Aug/17/2006 )

Dear all,

I would like to store my enzyme (metalloenzyme) in 25 - 50% glycerol, I have some question.

1. Does glycerol inteferes an enzymatic reaction if I use such enzyme in testing its kinetics (2-5 ul of the enzyme in 100 ul reaction) ?

2. If glycerol may intefere an enzymatic reaction (even in non-metalloenzyme), how is it interfere ?

Ps. the first question is more important than the second, and the second question is just ok if you answer in breif, please help me as much as you can.

Thanks a lot for your help.

Yes glycerol can interfere with enzymes, you have to dilute it enough. However 2.5µL in 100µL should be enough diluted.
enzymes for molecular biology are all in glycerol, and it works fine if you dilute them enough. (10 ot 20 times if I remember well)

I know New England Biolabs' restriction enzymes are supplied in 50% glycerol.

They recommend to keep the glycerol concentration at not greater than 5% during a reaction, meaning the restriction enzyme should not exceed 10% of the total reaction volume.

Thank you very much for all answers, : )