Methods
Summary
The activation or inhibition of cell-surface receptor, initiates a complex networking of downstream signaling cascades. Akt is implicated in regulating many biological processes, such as proliferation and cell growth. PI3K activation results in the activation of the Akt pathway by causing Akt to be phosphorylated, which is its 'active' state.
The HTRF® phospho Akt kit allows the measurement of the activation of PI3K because PI3K causes Akt to be phosphorylated and activated. We can measure levels of pAkt in response to different doses of our drugs. http://www.cisbio.com/drug-dis...
This experiment is relatively simple and has strong controls that can verify the results. It is also a quantitative method. That means that rather than a yes or no result, we will get a numerical score. This is very important because we need to be able to accurately describe how well each drug behaves and also what precise concentrations of each drug are most effective. Because the drugs will be tested in very similar conditions, often beside each other, we can also accurately describe if one drug is better than another.
Challenges
Our experts routinely undertake this experiment. This means that we do not expect any technical hurdles because these have been worked out in prior experiments. The main challenge in any experiment like this is rigor. Can you believe the results are genuine? Scientists overcome this hurdle by using 'controls'. We will include positive controls that we know should work and negative controls that we know shoudl have no effect. If these behave unusually, it tells us that something went wrong with the experiment. We can then fix that problem.
Pre Analysis Plan
Each individual drug will be tested over several concentrations and we will analyze this by creating a concentration curve that will show us what concentrations are effective. The lower the concentration, the better the drug will be, Indeed, a drug should have an effect at nanomolar concentrations. When small molecule need high doses, we often see side effects that make the drug unsafe. We will take all of the compounds and then compare them to see which drug is the most potent and has a range of useful doses that provides a full effect. We will also compare what the maximal effect of each drug is. Some drugs are only partially effective even when they are potent. 
Protocols
Browse the protocols that are part of the experimental methods.