Correlation of In-Cell Western Assays with Western Blots
Figure 1. In-Cell Western assays correlate well with Western blots.
Analysis of Akt phosphorylation
A) MCF-7 breast cancer cells were treated with recombinant human IGF-1. After fixation and permeabilization in microplate wells, phospho-Akt (S473) and total Akt protein levels were detected. Levels of phospho-Akt, normalized to total Akt protein levels in the same well, were quantified by analysis of fluorescence intensities. R&D Systems Phospho-Akt (S473) Cell-Based Infrared ELISA kit (KCBIR887) was used.
B) Phospho-Akt and total Akt were also detected by Western blot.
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- Kumar N et al. Multipathway model enables prediction of kinase inhibitor cross-talk effects on migration of Her2-overexpressing mammary epithelial cells. Mol Pharmacol 73:1668-1678 (2008).
1. Aguilar HN et al. PLoS ONE 5(4): e9965 (2010)