can someone please help biochem thank you The results from

can someone please help, biochem :/ thank you!!

The results from an enzyme purification procedure are shown below. The immunoprecipitation is carried out with an antibody known to bind the enzyme of interest. I run an SDS PAGE, and perform a Western blot with the same antibody used for immunoprecipitation, and see one single band at 40kD. However, when I run a native (non-denaturing) PAGE, followed by a Western blot with the same antibody, I see a single band of 160kD. What do you conclude from this experiment? Consider the specific activity table shown in the previous question. In measuring the activity of the enzyme after each purification step, what assumption/s of the Michaelis-Menten equation am I violating?

Solution

10. The specific activity suggests that the immunoprecipitation is a better method of purification as compared with the purification via salt precipitation.

SDS-PAGE results suggest that the enzyme is composed of four subunits of 40 KD molecular weight.