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How to confirm the role of akt in the signaling pathway by experiment results? It’s a question for many researchers and I hope I can introduce a way to solve the problem today. I would give an example to explain to way to prove our hypothesis. Assuming that we got a compound which can inhibit some kinase and it seems that the inhibition of this kinase would influence the cell proliferation, cell growth, or any other phenomenons through Akt signaling pathway, which kind of the experiment we should design to prove our presumption? How to confirm the relationship among different kinases which may be involved in this process? I would like to give a general idea and then give an example to help you understand the idea better. First, just use the inhibitor to block the targeted kinase. And we should test the level of proteins what we care about. Of course, the level of p-akt, the level of total Akt and level of akt downstream kinase and phosphorylated akt downstream kinase including GSK-3beta and p-GSK-3beta by western. In addition, don’t forget to take alpha-tubulin as the control. The we should collect other information or phenomenons in comparison with the level of kinases and p-kinases,for example, microscopic findings would be a good choice. But what you need depend on your experiment itself. It would be a good result if the microscopic findings can keep pace with the level of kinases in western blotting result. Then you can confirmed that AKt signaling pathway are really related to the disease or the targets you are interested in. But you can not be sure that the inhibitor block the target and induce the phenomenons we observed through the Akt signaling pathway. The you confirm whether the variation of level of p-akt is dose dependent to the variation of the level of the compound regardless of inducer existing. A dose-dependent manner can confirm that inhibition of that target by the compound can result in the phenomenon we observed through the regulating the activation of Akt. I wanna to take celecoxib, a cyclooxygenase-2 inhibitor, to explain what I mentioned above. As a Cyclooxygenase-2 inhibitor, celecoxib can reduce neointimal hyperplasia. But researchers wanted to show the role of Akt in this process. So they design a experiment. First, they culture cells with celecoxib and confirmed the phenomenon that neointimal hyperplasia reduced. Then they tested the level of Akt, p-Akt, GSK-3beta and p-GSK-3beta.(GSK-3beta is a downstream kinase of Akt). And they found that the level of p-akt and p-GSK-3beta both reduced while the level of total Akt remained the same level implying that celecoxib can reduce neointimal hyperplasia through inhibition of activity of Akt and GSK-3beta. Besides above, researchers used a gradient levels of PDGF to increase the activation of Akt.(PDGF is a growth factor which can induce the activation of Akt. In further, the higher level of PDGF existing, the more p-akt).And it confirmed that neointimal hyperplasia is reduced, when we cultured the cells with gradient celeoxib, is a dose-dependent manner with or without PDGF . p-akt, COX2 inhibitor, celeoxib And all the experiments together can prove that the Akt inhibition was in the process. It’s really a complex process to show the role of Akt. LY294002, nvp-bez235, high through-out screening Reference: [1] Yang, H. M., H. S. Kim, et al. (2004). ?Celecoxib, a cyclooxygenase-2 inhibitor, reduces neointimal hyperplasia through inhibition of Akt signaling.? Circulation 110(3): 301-308. Similar to The role of Akt in the process that neointimal hyperplasia is reduced by celeoxib Low concentration of NVP-BEZ235 could induce the Akt phosphorylation NVP-BEZ235 is a potent PI3K inhibitor. And it seems that NVP-BEZ235 should reduce the p-akt as PI3K, which can phosphorylate Akt, was blocked by NVP-BEZ235. High concentration ... The inhibition of bez235 on Akt and S6 In this poster, I want to give some details information about the function of BEZ235. I would list the assays related to BEZ235. And I ... BEZ235,a PI3K inhibitor, can target mutated p110-alpha NVP-BEZ235 is such an important PI3K inhibitor, and we would focus on the anti-tumor activity of BEZ235 in vivo in this poster. We all know ... We pays great attention to the purity, stability and activity of the products like Compound Libraries, compound library, High Throughput Screening, Screening Libraries, Screening Library & more. Not only could we provides the chemical test data, such as HNMR, LC-MS and HPLC, but also provides the reviews overall evaluations of the products from the customers who have used our products. Western Blot, MTT, RT-PCR, ICH photos or figures provide a double guarantee for the biological activities of our products in life-science research.
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