Radioenzymatc Assay - How to convert cpm values into enzyme specific activity (Oct/03/2008 )
Hi to all,
I am currently doing research on enzyme analysis. I am using C14 radioisotope as tracer to measure the amount of product formed from an enzyme extract that I partially purified from liver samples. I have prepared my radioisotope by mixing it to a known molar concentration of a similar but unlabeled molecule. I have counted the activity of the enzyme extract using liquid scintillation counter but I am facing challenges on how to actually use these CPM values to calculate for the activity of my enzyme. Pls help me with this. any information will be greatly appreciated. Thanks in advance...
Glenmer
-Glenmer-
QUOTE (Glenmer @ Oct 3 2008, 11:47 AM)
Hi to all,
I am currently doing research on enzyme analysis. I am using C14 radioisotope as tracer to measure the amount of product formed from an enzyme extract that I partially purified from liver samples. I have prepared my radioisotope by mixing it to a known molar concentration of a similar but unlabeled molecule. I have counted the activity of the enzyme extract using liquid scintillation counter but I am facing challenges on how to actually use these CPM values to calculate for the activity of my enzyme. Pls help me with this. any information will be greatly appreciated. Thanks in advance...
Glenmer
I am currently doing research on enzyme analysis. I am using C14 radioisotope as tracer to measure the amount of product formed from an enzyme extract that I partially purified from liver samples. I have prepared my radioisotope by mixing it to a known molar concentration of a similar but unlabeled molecule. I have counted the activity of the enzyme extract using liquid scintillation counter but I am facing challenges on how to actually use these CPM values to calculate for the activity of my enzyme. Pls help me with this. any information will be greatly appreciated. Thanks in advance...
Glenmer
you must know the specific activity of your radioactive probe which means the ratio of radioactive probe to total probe; then you can easily convert cpm (or even better dpm) into catalyzed probe;
commonly you take mg protein and time in min into account
-The Bearer-