I have obtained concentrations for 3 unknown proteins through BCA assay. I ran kinetics on a spectrophotometer for each unknown at two different different volumes(5uL and 10uL). We only did 1 trial for each volume for each unknown(6 measurements). We ran out of time to do more and we were having some issues with the substrates. Regardless, I now have to write a report on the experiment.

 

This was a citrate synthase experiment. I have no idea what I am looking for. What I do know is that when we increase the volume of the protein samples the ABS/min goes up. Im assuming I should solve for Km and Vmax, but other than that I have no idea what this means as it relates to citrate synthase. Ive asked the instructor and am told to interpret it myself. I am lost here. Help please.

 

 

I have obtained concentrations for 3 unknown proteins through BCA assay. I ran kinetics on a spectrophotometer for each unknown at two different different volumes(5uL and 10uL). We only did 1 trial for each volume for each unknown(6 measurements). We ran out of time to do more and we were having some issues with the substrates. Regardless, I now have to write a report on the experiment.

 

This was a citrate synthase experiment. I have no idea what I am looking for. What I do know is that when we increase the volume of the protein samples the ABS/min goes up. Im assuming I should solve for Km and Vmax, but other than that I have no idea what this means as it relates to citrate synthase. Ive asked the instructor and am told to interpret it myself. I am lost here. Help please.

 

 

You mean three different concentrations of different proteins, or three different concentrations of the same protein?

 

You added Ellman's reagent (DTNB) and citrate synthase (your protein?), and measured the absorbance of the formation of the product of the reaction (TNB) at 412 nm?