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I have 100 micrograms of STAT1 and its molecular weight is 90 kDa. I calculated the number of moles to be 1 nmoles. The specific activity of JAK1 (the kinase) is 110 nmoles of phosphate per minute per mg of total protein at 30°C and the activity was determined at a final concentration of 0.83 micrograms/milliliters. What amout of JAK1 do I need?

I tried many ways but I don't get how to link the data.

Thank you.

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    $\begingroup$ How many phosphorylation sites does STAT1 have? How long to you want to wait for the reaction to be done? $\endgroup$ – Karsten Theis May 1 at 0:48
  • $\begingroup$ Only one site Tyr701, I'm not sure about the amount of time of the reaction. $\endgroup$ – Felipe May 1 at 0:51
  • $\begingroup$ "I have 100 micrograms of STAT1 and its molecular weight is 90 kDa. I calculated the number of moles to be 1 nmoles. " Calculate again. $\endgroup$ – imalipusram May 1 at 1:05
  • $\begingroup$ It would take 1 minute to phosphorylate 0.11 nmoles of substrate if you add a microgram of enzyme (in a reaction volume of 1 mL). If you have 1 nmole substrate instead, it would take about 10 minutes with the same amount of enzyme. $\endgroup$ – Karsten Theis May 1 at 1:07
  • $\begingroup$ Thank you! How did you get to the 1 minute___0.11 nmole relation? $\endgroup$ – Felipe May 1 at 1:29

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