Protein Expression in Cells

Protein Expression Athena application applied for detection of cell viability (Autophagy), protein expression and protein phosphorylation Assay in breast cancer research:

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[Neratinib + Valproate] exposure permanently reduces ERBB1 and RAS expression in 4T1 mammary tumors and enhances M1 macrophage infiltration.

Booth et al., Oncotarget, 26;9(5):6062-6074, Dec 2017

Breast cancer drug neratinib has been found to block the function of the Ras genes which are among the first to be linked to cancer development.

Image analysis showed Neratinib down-regulating the expression of mutant K-RAS and mutant N-RAS.

Neratinib and HDAC inhibitors interact to reduce the expression of mutant N-RAS and mutant K-RAS. Spiky and PANC1 cells that express a mutant N-RAS and a mutant K-RAS, respectively, were treated with vehicle control and neratinib (50 nM). The cells were fixed in place and immunostaining performed with DAPI counter-stain to determine the expression levels and cellular localization of N-RAS and K-RAS at 60× magnification using Hermes HCS system.

“In-Cell Westerns” Assay

A novel approach to detect gross changes in protein expression and protein phosphorylation

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Rationally repurposing Ruxolitinib (Jakafi®) as a solid tumor therapeutic

Tavallai Mehrad, Booth Laurence, Roberts Jane L, Poklepovic Andrew, Dent Paul ; Frontiers in Oncology ,vol.6,2016 

Assessments of total protein expression or post translational modifications, such as phosphorylation, can be made via fluorescence intensity measurements.  Such “in-cell western” analysis uses unbiased pre-programmed electronic data acquisition, much as has previously been performed for the last 45 years using SDS-PAGE and western immunoblotting.

The table above compares main differences between the conventional western blot assay and protein expression endearments using the Hermes high content imaging system.