The In-Cell Western assay was used to measure effects of the MEK inhibitor U0126 on EGFR-induced signaling.
Timing/Kinetics of Signal Transduction
The kinetics of cellular β-catenin accumulation upon stimulation with Wnt3a was studied.
EC50 and IC50 Determinations
In-Cell Western assays were used to determine EC50 and IC50.
Tau Protein Accumulation and Inhibition
The In-Cell Western Assay is a powerful technique for virologists to define specific viral characteristics, such as viral infectivity and mechanism of action, or to screen antiviral therapeutics and test vaccine efficacy. Results from the In-Cell Western have been shown to correlate well with other virology methods and demonstrate that the In-Cell Western is time efficient and well suited for the study of both cytopathic and non-cytopathic viruses.
For instance, when compared with three common methods to quantify Hantaan virus (HTNV) replication: quantitative reverse transcription polymerase chain reaction (qRT-PCR), flow cytometry (FCM), and enzyme-linked immunosorbent assay (ELISA), the In-Cell Western Assay minimized operator subjectivity and was found to have results that were superimposable with results from the other assays.6 This study highlights the capability of the assay for assessing HTNV replication.
For more details about using the In-Cell Western in virology, visit the Virology application page.
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Fabiani, M., et. al. (Fig. 11) found that the In-Cell Western Assay and standard plaque assay (SPA) produced nearly superimposable results. However, the In-Cell Western Assay was faster to detect viral infectivity, enabled many samples to be measured in parallel, and was found to be more suitable for titrations that produced plaques too numerous to count using SPA.7
Cell Surface Proteins
On-Cell Western Assays are a variation of the In-Cell Western Assay in which the cells are not permeabilized. For On-Cell Westerns, the unpermeabilized cells are stained with antibodies against extracellular protein domains so only cell surface antigens are detected. Using On-Cell Western Assays, you can quantitatively monitor cell surface protein expression.