Enzyme activity - Enzyme activity (Feb/14/2006 )
hi there
I am doing NQO1 enzymatic activity Dicumarol as an inhibitor. I am using UV-Vis spectrophotometer from Agilent rechnologies. My problem is that i am getting the negative reaction rate after adding the Dicumarol in the reaction mixture. So I want to know whether it is right or wrong. I am waiting passionately for your replies. Please,help me.
If you can please attach the detailed protocol and the calculation method in the protocol online. May it will be helpful for others also.
Thanking you in advance.
well your post is not very clear, but assumoing dicumarl is the inhibitor, you're locking the reaction. I don't understand very well the "negative reaction rate" term
May you clarify your exp?
May you clarify your exp?
Well,,thanx for ur interest. I mean that i am using the uv-vis chemstation software,,so it gives the reaction rate after the calculation of the absorbance . So when i add only enzyme n substrate the rate value is positive and in the next step to the same new sample after i added the dicumarol and then substrate then the values r in the negative sign,,,so i am bit confused.the graphical representation is also different ,,,without dic it was increasing over time but with dic addition it was decreasing,,,,,so what do you think ???what shud i do???
what are the compounds of your exp? Maybe the thing that absob the loght degrades itself...
i am using cytochrome c,,NADH,Menadione and Dicumarol in the rxn mixture.so please,how can u help me???
thanking u in advance.
hi:
Since you got a negative reaction rate, there must be other side-reaction occured. Do you use phosphate buffering system? It may cause it. Because you use spectrotometry, single reaction in the wave-length rage was first suggested to be confirmed . Regards