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| | PI3K Inhibition Assay |
| Description: | PI3K inhibition assay (PI3K Assay (Emmanuelle M, Huang Y, Yan H G et al. Targeting Protein Translation in Human Non-Small Cell Lung Cancer via Combined MEK and Mammalian Target of Rapamycin Suppression. Cancer Res 67:(23). (2007).) was carried out by PI3 Kinase activity/inhibitor assay kit, where PI3 kinase reaction was set up in Glutathione-coated strips/plate for inhibitor reaction. Kinase and inhibitors were pre-incubated for 10 minutes prior to the addition of PIP2 substrate. 5 uL of 5x kinase reaction buffer were added in each well followed by the further addition of 5 uL/well of PIP2 substrate. Then distilled H2O was added to each well so as to make up a final volume of 25 uL/well. Incubation was done at rt for 1 hour which was followed by washing the wells 3 times with 200 uL of 1xTBST per well and then 2 times with 200 uL of IX TBS per well. Then 100 uL of the Substrate TMB per well was added and then to keep for colour development in the dark. |
| Affinity data for this assay | |
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